Team:Gothenburg-Sweden/test1

From 2010.igem.org

(Difference between revisions)
 
(183 intermediate revisions not shown)
Line 1: Line 1:
<html xmlns="http://www.w3.org/1999/xhtml">
<html xmlns="http://www.w3.org/1999/xhtml">
 +
<head>
 +
<meta http-equiv="content-type" content="text/html; charset=utf-8" />
 +
<title>Chalmers University of Technology</title>
 +
<style type="text/css">
<style type="text/css">
-
/* Wiki Hacks - START */
 
-
/* Author: Pieter van Boheemen */
 
-
/* Team: TU Delft */
 
-
h2 {background:none; text-decoration:none;}
 
-
#globalWrapper { background-color: transparent; border: none; margin: 0; padding: 0;}
 
-
#content { background-color: transparent; border: none; padding: 0; margin: 0; width: 100%;}
 
-
#bodyContent { border: none; padding:0; margin:0; width:100%;}
 
-
#top-section { height: 20px; margin-top: -5px; margin-left: auto; margin-right: auto; margin-bottom: 0 !important; margin-bottom: 10px; padding:0; border: none;}
+
.style4 {
-
#p-logo { height:1px; overflow:hidden; display: none;}
+
font-size: 11px;
-
#search-controls { overflow:hidden; display:block; background: none; position: absolute; top: 100px; right: 40px;}
+
font-weight: bold;
-
.left-menu { width: 400px; display:block; float:left; margin-top:-80px; border: none;}
+
}
-
.left-menu ul { border: none;}
+
.style5 {font-size: 11px}
-
#menubar.right-menu { width:300px; display:block; float:left; margin-top:-80px; border: none;}
+
.style9 {
-
.right-menu ul { border: none;}
+
font-size: 12px;
-
#footer-box { background-color: transparent; border: none; width: 965px; margin: 0 auto; padding: 0;}
+
color: #00456e;
-
#footer { border: none; width: 1300px; margin: 0; padding: 0;}
+
font-weight: bold;
-
.firstHeading { display: none;}
+
}
-
#f-list a { color: #333; font-size: 10px;}
+
.style11 {font-size: 10px; color: #00456e; }
-
#f-list a:hover { color: #666;}
+
 
-
#footer ul { margin: 0; padding: 0;}
+
#myTitle{
-
#footer ul li { margin-top: 0; margin-bottom: 0; margin-left: 10px; margin-right: 10px; padding: 0;}
+
    font-family: Arial, Helvetica, sans-serif;
-
/* Wiki Hacks - END */
+
    font-weight:normal;
 +
    color: #00456e;
 +
    font-size: 16px;
 +
    height: 36px;
 +
   
 +
    padding-top: 10px;
 +
    padding-right:-200px;
 +
    margin-left: 20px;
 +
    text-align:center;
 +
    background: url(https://static.igem.org/mediawiki/2010/6/64/Title_center.png) left top;
 +
    background-repeat:no-repeat;
 +
 
 +
}
 +
 
 +
/*to be modified */
#contentSub {
#contentSub {
display:none;
display:none;
}
}
 +
#siteSub {
#siteSub {
-
display:none;
+
display:none;padding:-200px;margin:0;
}
}
-
#search-controls {
 
-
display:none;
 
-
}
 
.firstHeading {
.firstHeading {
-
display:none;
+
display:none;padding:0;margin:0;
}
}
#search-controls {
#search-controls {
-
margin-top:30px;
+
padding-top:10px;
}
}
-
#footer-box {
+
 
-
display:none;
+
 
 +
form {
 +
margin: 0px;
 +
padding: 0px;
}
}
 +
 +
h2 {background:transparent; text-decoration:none;  }
 +
#globalWrapper { background-color: transparent; border: none; margin-bottom: 0px; padding: 0;}
 +
#content { background-color: transparent; border: none; padding: 0; margin-bottom: 0px; width: 100%;}
 +
#bodyContent { border: none; padding:0; margin-bottom:0px; width:100%;}
 +
 +
#top-section { background-color: transparent; height: 20px; margin-top: -30px; margin-left: auto; margin-right: auto; margin-bottom: 0 !important; margin-bottom: 0; padding:0; border: none; }
 +
#p-logo {  display: none;}
 +
#search-controls { display:none;}
 +
 +
#menubar.left-menu {width: 400px;  display:block; float:left; margin-top:-50px; border: none;}
 +
#menubar.left-menu ul { border: none;}
 +
#menubar.left-menu a{color:#5698b0;}
 +
#menubar.left-menu a:hover{color:#e6f6fb;}
 +
#menubar.left-menu a:selected{color:#e6f6fb;}
 +
 +
#menubar.right-menu {  width:300px; display:block; float:left; margin-top:-50px; border: none;}
 +
#menubar.right-menu ul {  border: none;}
 +
#menubar.right-menu a{color:#5698b0;}
 +
#menubar.right-menu a:hover{color:#e6f6fb;}
 +
 +
 +
/* Wiki Hacks - END */
 +
 +
 +
 +
 +
/*to be modified */
 +
 +
body {background:url(https://static.igem.org/mediawiki/2010/8/8e/Shadow.png) repeat-y top center;
 +
  padding: 0 0 0 0;
 +
 
 +
    }
a{
a{
     font-family: Arial, Helvetica, sans-serif;
     font-family: Arial, Helvetica, sans-serif;
Line 56: Line 101:
     color: #06639e;
     color: #06639e;
     text-decoration:none;
     text-decoration:none;
 +
    padding:0;
 +
    margin:0;
}
}
-
p {
 
-
font-size:12px;
 
-
}
 
-
p2{font-size:11px;
 
-
color:#07374E;
 
-
 
-
}
 
a:hover{
a:hover{
 +
padding:0;margin:0;
     text-decoration: underline;
     text-decoration: underline;
     color: #000066;
     color: #000066;
 +
}
}
 +
a:visited{color:#06639E;}
a:visited{color:#06639E;}
Line 79: Line 122:
}
}
-
body
 
-
{
 
-
    font: 12px Arial, Helvetica, sans-serif;
 
-
    color: #000000;
 
-
    line-height: 20px;
 
-
    background: #FEFEFE url(https://static.igem.org/mediawiki/2010/5/5b/BgGo.jpg);
 
-
    background-repeat:repeat-x;
 
-
    background-position: top;
 
-
}
 
-
ul{list-style:none;}
+
#all{
 +
  padding:0;
 +
  margin:0;
 +
  padding-left:10%;
 +
  background:  url(https://static.igem.org/mediawiki/2010/3/3a/Bg.png) top center repeat-y;
 +
}  
-
#mainbg{
 
-
    width: 1016px;
 
-
    margin: 0 auto;
 
-
    padding-top:20px;
 
-
}
 
-
 
-
#main{
 
-
    margin: 0 auto;
 
-
    width: 1016px;
 
-
    padding-top: 30px;
 
-
}
 
#header{
#header{
-
    height: 295px;
+
  height: 290px;
-
    width: 100%;
+
  background:url(https://static.igem.org/mediawiki/2010/5/50/BannerGo.png) no-repeat;  
-
padding-left:85px;
+
  margin-left:15px;
-
}
+
-
 
+
}
-
#content{
+
#search {height: 30px;}
 +
#myContent{
 +
margin-top: 0px;
}
}
 +
 +
/* left layer starts */
#left{
#left{
-
     width:180px;
+
     width:138px;
-
 
+
    padding-left:45px;
     float: left;
     float: left;
-
 
 
}
}
Line 125: Line 156:
     font-family: Comic Sans;
     font-family: Comic Sans;
     font-weight:600;
     font-weight:600;
-
     color: #ffffff;
+
     color: #fff;
     font-size: 16px;
     font-size: 16px;
     text-align:center;
     text-align:center;
     width:100%;
     width:100%;
     height: 30px;
     height: 30px;
-
     padding-top: 9px;
+
     padding-top: 7px;
-
     background: url(https://static.igem.org/mediawiki/2010/6/60/Title_left.png);
+
    margin-top:7px;
 +
     background: url(https://static.igem.org/mediawiki/2010/a/aa/CatTitle.png);
     background-repeat:no-repeat;
     background-repeat:no-repeat;
     background-position:top left;
     background-position:top left;
}
}
-
#left .company{
+
.left_grad{
-
     padding-top: 10px;
+
   
-
font-size:11px;
+
    background-position: top left;
 +
     padding-top: 0px;
 +
    margin-top: 0px;
}
}
-
.numleft{
+
.categories{
-
     background: url(https://static.igem.org/mediawiki/2010/0/0a/Numbg.png);
+
     padding-top:0px;
-
    background-repeat: no-repeat;
+
background: url(https://static.igem.org/mediawiki/2010/1/10/CatCon.png) top repeat-y;
-
    background-position: left top;
+
     min-height:100px;
-
    float:left;
+
     border: 1px solid #0485B0;
-
    width: 37px;
+
-
     min-height: 37px;
+
-
     font-family: Arial,Helvetica,sans-serif;
+
-
 
+
-
    font-weight: bold;
+
-
    color: #ffffff;
+
-
    text-align: center;
+
-
    margin: 0px 8px 0px 8px;
+
}
}
-
.numleft p{
+
.categories a{
-
     padding-top: 2px;
+
     font-family: Arial, Helvetica, sans-serif;
-
    font-size: 16px;
+
-
}
+
-
 
+
-
 
+
-
.newsright{
+
-
    padding-left: 53px;
+
-
    padding-right: 10px;
+
-
+
-
}
+
-
 
+
-
#left .read a{
+
-
    color: #06639e;
+
     font-weight:normal;
     font-weight:normal;
-
     font-style: italic;
+
     color: #0485B0;
-
     font-size: 12px;
+
     text-decoration: none;
-
     text-decoration:none;
+
     text-align:left;
-
     border-bottom: 1px #06639e;
+
     padding: 6px 0px 6px 20px;
-
     margin-right: 10px;
+
    font-size:12px;
 +
     padding-top:5px;
 +
    display:block;
 +
 
}
}
-
#left .read a:hover{border-bottom: 1px dotted #06639e;}
+
.categories a:hover{
 +
  text-decoration:none;
-
.left_grad{
+
  font-size:12px;
-
    background: url(https://static.igem.org/mediawiki/2010/d/d0/Left_grad.png) repeat-y;
+
  font-weight:700;
-
    background-position: top left;
+
  color: #fff;
-
    padding-top: 0px;
+
  background:url(https://static.igem.org/mediawiki/2010/4/47/CatAhover.png);
}
}
 +
/* left layer ends */
-
.list_border{
+
/* center layer starts */
-
    background: url(https://static.igem.org/mediawiki/2010/d/d0/Left_grad.png) repeat-y;
+
-
    background-position:left top;
+
-
    height: 5px;
+
-
    margin-bottom: 10px;
+
-
}
+
#center{
#center{
-
     width: 616px;
+
     width: 600px;
-
     padding-left: 10px;
+
     padding-left: 20px;
     float: left;
     float: left;
}
}
-
 
#center p{
#center p{
     padding-left:0px;
     padding-left:0px;
     padding-right:0px;
     padding-right:0px;
     padding-bottom: 10px;
     padding-bottom: 10px;
-
color: #07374E;
+
-
 
+
}
}
 +
    /* menu starts */
-
.text{
+
     
-
    padding: 10px;
+
        #Title2{
-
    background: #ffffff url(https://static.igem.org/mediawiki/2010/d/da/Text_bg.png);
+
 
-
    background-position: bottom;
+
          height: 36px;
-
    background-repeat: repeat-x;
+
          width: 600px;
-
    min-height: 64px;
+
          padding-top: 7px;
-
+
          padding-bottom:5px; 
-
}
+
          background: url(https://static.igem.org/mediawiki/2010/5/50/Title_center1.png);
 +
          background-repeat:no-repeat;
 +
          background-position: center;
 +
          }
-
.lab{
+
#sddm
-
  padding-top:10px;
+
{
 +
 
 +
  padding-left:62px;
 +
  padding-top: 0px;
 +
}
 +
 +
#sddm li
 +
{     
 +
list-style: none;
 +
float:left;
 +
text-align:center;
 +
}
 +
 +
 +
#sddm li a
 +
 +
{ display: block;
 +
margin: 0 0 0 0;
 +
padding: 5px 0px;
 +
width: 96px;
 +
height:27px;
 +
text-align: center;
 +
text-decoration: none;
 +
font:  14px Rockwell ;
 +
font-weight:500;
 +
 +
}
 +
#active {
 +
         
 +
 +
}
 +
#active a{
 +
                          background: url(https://static.igem.org/mediawiki/2010/0/0c/MActive2.png) top;
 +
  color:#FFFFFF;
 +
  font: 13px arial;
 +
          font-weight:600;
 +
                     
 +
}
 +
#sddm li a:hover
 +
{ background: url(https://static.igem.org/mediawiki/2010/5/5e/MaHover.png) top center no-repeat;
 +
color:#fff;
 +
font:  13px arial;
 +
font-weight:600;
 +
}
 +
 +
 +
#sddm div
 +
{ position: absolute;
 +
visibility: hidden;
 +
margin-top: -8px;
 +
                     
 +
padding: 0px;
 +
background: #fff ;
 +
border: 1px solid #9CD2ED ;
 +
}
 +
 +
 +
#sddm div a
 +
{ position: relative;
 +
height:16px;
 +
display: block;
 +
margin: 0px;
 +
padding: 4px 20px;
 +
                                padding-top:auto;
 +
                                padding-bottom:auto;
 +
width:auto;
 +
min-width: 100px;
 +
font-family: sans-serif, Tahoma, Verdana, Geneva, Arial, Helvetica;
 +
text-align: left;
 +
text-decoration: none;
 +
background: transparent;
 +
color: #2e6d95;
 +
font-size: 11px ;
 +
border:none;
 +
 +
}
 +
 +
#sddm div a:hover
 +
{ background: #61b5da  url(https://static.igem.org/mediawiki/2010/9/9d/MArrow.png) no-repeat;
 +
 +
color: #FFF;
 +
text-decoration:none;
 +
font:  11px arial;
 +
font-weight:500;
 +
}
 +
 +
/* menu ends */
 +
 +
#text{
 +
    text-align:left;
 +
width:100%;
 +
margin: 0px 10px 10px 20px;
 +
    background: #fff ;
}
}
-
#Title2{
+
#text p
-
    font-family: Arial, Helvetica, sans-serif;
+
{
-
    font-weight:normal;
+
  padding: 10px 10px 10px 25px;
-
    color: #00456e;
+
  font-family: "Times New Roman", Times, serif;
-
    font-size: 16px;
+
  font-size:13px;
-
    height: 36px;
+
  line-height:19px;
-
    width: 616px;
+
-
    padding-top: 10px;
+
-
text-align:center;
+
-
    background: url(https://static.igem.org/mediawiki/2010/6/64/Title_center.png);
+
-
    background-repeat:no-repeat;
+
-
    background-position: top;
+
}
}
 +
 +
.logo{
 +
float:left;
 +
padding: 10px 20px 10px 20px;
 +
}
 +
 +
#text p{color: #07374E;}
.read{
.read{
     text-align: right;
     text-align: right;
     padding-bottom:5px;
     padding-bottom:5px;
-
    padding-right: 5px;
+
  /* padding-right: 5px;*/
}
}
Line 248: Line 357:
     height: 26px;
     height: 26px;
     padding-top: 3px;
     padding-top: 3px;
-
     background: url(https://static.igem.org/mediawiki/2010/0/07/Read.png);
+
     background: url(images/read.png);
     background-repeat: no-repeat;
     background-repeat: no-repeat;
     background-position: top;
     background-position: top;
Line 258: Line 367:
     font-style: italic;
     font-style: italic;
     font-size: 12px;
     font-size: 12px;
-
    padding-right: 15px;
+
  /* padding-right: 15px;*/
     text-decoration:none;
     text-decoration:none;
}
}
-
.read a:hover{
+
#team {
-
    text-decoration:underline;
+
padding: 20px 10px 20px 30px;
 +
 
}
}
 +
 +
/* center layer ends */
 +
 +
/* right layer starts */
#right{
#right{
-
     width:180px;
+
     width:160px;
-
     padding-right: 19px;
+
     padding-left: 20px;
-
     float:right;
+
    /*padding-right: 135px;*/
 +
     float:left;
}
}
-
#right h2 {
+
#right .read a{
 +
   
 +
    color: #06639e;
 +
    font-weight:normal;
 +
    font-style: italic;
 +
    font-size: 12px;
 +
    text-decoration:none;
 +
    border-bottom: 1px #06639e;
 +
    padding-right:10px;
 +
}
 +
 
 +
#right .read a:hover{border-bottom: 1px dotted #06639e;}
 +
 
 +
 
 +
#lab{
 +
 +
padding-top:0px;
 +
background: url(https://static.igem.org/mediawiki/2010/1/10/CatCon.png) top left repeat-y;
 +
min-height:100px;
 +
/*margin-left:30px;*/
 +
border: 1px dotted #0485B0;
 +
}
 +
.numleft{
 +
background: url(https://static.igem.org/mediawiki/2010/2/22/Numbg1.png);
 +
background-repeat: no-repeat;
 +
background-position: left top;
 +
float:right;
 +
width: 37px;
 +
min-height: 37px;
 +
font-family: Arial,Helvetica,sans-serif;
 +
 +
font-weight: bold;
 +
color: #ffffff;
 +
text-align: center;
 +
margin: 25px 8px 0px 8px;
 +
}
 +
 +
 +
.numleft p{
 +
padding-top: 2px;
 +
                          padding-left: 1px;
 +
font-size: 16px;
 +
 +
}
 +
 +
 +
.newsright{
 +
    text-align:left;
 +
margin: 10px 50px 0 10px;
 +
}
 +
 +
.newsright p{
 +
font-size:11px;
 +
            color:#07374E;
 +
            margin-top:7px;
 +
 +
}
 +
 
 +
.Title3 {
     font-family: Comic Sans;
     font-family: Comic Sans;
     font-weight:600;
     font-weight:600;
-
     color: #ffffff;
+
     color: #fff;
     font-size: 16px;
     font-size: 16px;
     text-align:center;
     text-align:center;
 +
    width:100%;
     height: 30px;
     height: 30px;
-
    padding-top: 9px;
+
    padding-top: 7px;
-
     background: url(https://static.igem.org/mediawiki/2010/6/60/Title_left.png);
+
    margin-top:7px;
 +
margin-left:0px;
 +
     background: url(https://static.igem.org/mediawiki/2010/c/cf/CatTitle1.png) ;
     background-repeat:no-repeat;
     background-repeat:no-repeat;
     background-position:top left;
     background-position:top left;
}
}
-
.categories{
 
-
    min-height:100px;
 
-
}
 
-
.categories a{
+
/* right layer ends */
-
    font-family: Arial, Helvetica, sans-serif;
+
-
    font-weight:normal;
+
-
    color: #07374E;
+
-
    text-decoration: none;
+
-
    font-size:11px;
+
-
}
+
-
.categories a:hover{
 
-
    background-color:#2286C9;
 
-
color:#FFFFFF;
 
-
}
 
-
.categories ul{
+
/* footer layer starts */
-
    padding-left: 5px;
+
#myfooter
-
    padding-right: 5px;
+
-
}
+
-
 
+
-
.categories ul li{
+
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    <head>
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        <title>Chalmers</title>
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                              <div class="Title1">Team</div>
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                                  <div class="categories">
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 +
                                          <a href="#">Introduction</a>
 +
                                          <a href="#">CHALMERS</a></li>
 +
                                          <a href="#">Supervisors</a></li>
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                                          <a href="#">Students</a>
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                              <div class="Title1">FUSS</div>
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 +
                                            <a href="#">What's all about?</a>
 +
                                            <a href="#">Background</a>
 +
                                            <a href="#">Protein Fusion</a>
 +
                                            <a href="#"> Methods</a>
 +
                                            <a href="#">Preliminary Results</a>
 +
                                       
 +
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<div id="Title2"><ul id="sddm">
     <li><a href="https://2010.igem.org/Team:Gothenburg-Sweden">Home</a></li>
     <li><a href="https://2010.igem.org/Team:Gothenburg-Sweden">Home</a></li>
     <li><a href="https://2010.igem.org/Team:Gothenburg-Sweden/Project"  
     <li><a href="https://2010.igem.org/Team:Gothenburg-Sweden/Project"  
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     <li><a href="https://2010.igem.org/Team:Gothenburg-Sweden/Sponsors">Sponsors</a></li>
     <li><a href="https://2010.igem.org/Team:Gothenburg-Sweden/Sponsors">Sponsors</a></li>
-
     <li><a href="https://2010.igem.org/Team:Gothenburg-Sweden/Contact">Contact Us</a></li>
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<p> We are a team of 8 students from Chalmers University of Technology who  will represent Gothenburg, SWEDEN in this year’s iGEM competition. We  have started with a promising idea that combines the cutting edge  technologies available in the field of Synthetic Biology. Our research  basically aims to constructing a reporter mechanism for cellular stress  in yeast by tagging protein and peptides affiliated with the stress  activated SNF1 complex with fluorescent markers. </p>
 +
                 
 +
         
 +
 +
 +
                            <div id="myTitle">Chalmers University of Technology  </div>  
 +
                        <div class="logo"> <img src="https://static.igem.org/mediawiki/2010/a/a5/Chalmers_c_logo.jpg" width="133" height="152" align="left"></div>
 +
                   
 +
                              <p>
 +
                                <strong>Chalmers in central Göteborg on the Swedish West Coast</strong></p>
 +
                       
 +
                              <p>Chalmers is a university of technology in which research and  teaching  are conducted on a broad front within technology, natural  science and  architecture. Our inspiration lies in the joy of discovery  and the  desire to learn. Underlying everything we do is a wish to  contribute to  sustainable development both in Sweden and world-wide.<br>
 +
                                  <br>
 +
                                Our  research work deals ultimately with improving people’s  conditions, and  we often cross traditional boundaries in order to  solve the problems of  the future. Chalmers has become strong within  several areas of science,  and some of our research leads its field  internationally. We wish in  particular to develop and strengthen our  research in the fields of  bioscience, information technology,  environmental science and  nanotechnology.<br>
 +
  <br>
 +
                                Chalmers is a pioneer in Europe in the field of  start-up  companies, and has built up an efficient system of innovation  in order  to find applications in research into the formation of  companies. <br>
 +
  <br>
 +
                                Chalmers has great aspirations to develop continually  by  opening doors to the outside world and by thinking innovatively. We    have a close and productive collaboration with business and society at    large. In this way we reinforce our own development, at the same time  as  we contribute to the growth of society.<br>
 +
  <br>
 +
                                Chalmers was founded in  1829 as a result of a donation from  the director of the Swedish East  India Company, William Chalmers. His  motto, and ours, is:<br>
 +
  <br>
 +
                                Avancez!</p>
 +
                 
 +
                         
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+
                       
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                <!-- header ends -->
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-
                                        <ul>
+
-
                                            <li><a href="#">What's all about?</a></li>
+
-
                                            <li><a href="#">Theoretical Background</a></li>
+
-
                                            <li><a href="#">Fusion Protein Design</a></li>
+
-
                                            <li><a href="#">Experimental Methods</a></li>
+
-
                                            <li><a href="#">Preliminary Results</a></li>
+
-
                                        </ul>
+
-
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+
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+
-
    <ul>
+
-
                                            <li><a href="#">Introduction</a> </li>
+
-
<li><a href="#">Chalmers University of Technology</a></li>
+
-
                                            <li><a href="#">Supervisors</a></li>
+
-
                                            <li><a href="#">Students</a></li>
+
-
                                         
+
-
                                         
+
-
                                        </ul>
+
-
 
+
-
                                  </div> 
+
-
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+
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                              </div> 
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                                      <tr>
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                                        <td width="145" rowspan="2" valign="top"><span class="style8"><img src="https://static.igem.org/mediawiki/2010/e/eb/Goutham.jpg" alt="" width="133" height="200" class="img1" /></span></td>
 +
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 +
                                              <td><span class="style9">Goutham Vemuri</span></td>
 +
                                            </tr>
 +
                                            <tr>
 +
                                              <td><span class="style11">supervisor</span></td>
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                                        </table></td>
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                                        <td valign="top"><p>&nbsp;</p>
 +
                                        </td>
 +
                                        <td width="214" valign="top"><p class="style5">&nbsp;</p>
 +
                                  </table>
 +
<p>&nbsp;</p>
 +
<table width="560" border="1">
 +
                                      <tr>
 +
                                        <td width="145" rowspan="2" valign="top"><span class="style8"><img src="https://static.igem.org/mediawiki/2010/3/32/Per.jpg" alt="" width="133" height="200" class="img1" /></span></td>
 +
                                        <td width="174" height="59" valign="top"><table width="150" border="5" cellpadding="0" cellspacing="0">
 +
                                            <tr>
 +
                                              <td><span class="style9">Per Sunnerhagen </span></td>
 +
                                            </tr>
 +
                                            <tr>
 +
                                              <td><span class="style11">supervisor</span></td>
 +
                                            </tr>
 +
                                        </table></td>
 +
                                      </tr>
 +
                                      <tr>
 +
                                        <td valign="top"><p>&nbsp;</p></td>
 +
                                        <td width="214" valign="top"><p class="style5">&nbsp;</p>
 +
                                  </table>
 +
<p>&nbsp;</p>
 +
<table width="560" border="1">
 +
                                      <tr>
 +
                                        <td width="145" rowspan="2" valign="top"><span class="style8"><img src="https://static.igem.org/mediawiki/2010/8/88/Markus.jpg" alt="" width="133" height="200" class="img1" /></span></td>
 +
                                        <td width="169" height="59" valign="top"><table width="172" border="5" cellpadding="0" cellspacing="0">
 +
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 +
                                              <td width="136"><span class="style9">Marcus Wilhelmsson  </span></td>
 +
                                            </tr>
 +
                                            <tr>
 +
                                              <td><span class="style11">supervisor</span></td>
 +
                                            </tr>
 +
                                        </table></td>
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                                      </tr>
 +
                                      <tr>
 +
                                        <td valign="top"><p>&nbsp;</p></td>
 +
                                        <td width="219" valign="top"><p class="style5">&nbsp;</p>
 +
                                  </table>
 +
  <td>&nbsp;</td>
 +
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 +
<tr></tr>
 +
<tr>
 +
  <td>&nbsp;</td>
 +
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 +
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 +
                       
 +
                     
 +
 +
               
 +
                            <div id="myTitle">Students</div>
                              
                              
-
                        </div>
+
                              <div id="team">
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-
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+
                                      <td width="151" height="59" valign="top"><table width="150" border="5" cellpadding="0" cellspacing="0">
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                                     <div class="categories">
+
                                            <td><span class="style9">Malin Linden</span></td>
-
                                        <ul>
+
                                          </tr>
-
                                             <li><a href="#">Göteborgs-Posten to publish...</a></li>
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                                          <tr>
-
                                            <li><a href="#">Team members reunite...</a></li>
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                                            <td><span class="style11">student</span></td>
-
                                            <li><a href="#">Sponsor deal set...</a></li>
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                                          </tr>
-
                                            <li><a href="#">Yeast resistance for sporulation... </a></li>
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                                      </table></td>
-
                                       
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                                      <td width="237">&nbsp;</td>
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                                         </ul>
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                                     </tr>
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                                     </div>
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                                    <tr>
-
                           
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                                      <td colspan="2" valign="top"><table width="363" height="150" border="1">
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                          </div>
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                                          <tr>
-
                       
+
                                            <td width="147"><span class="style4">Age:</span></td>
-
                            <div class="list_border"></div>  
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                                            <td width="158"><span class="style5">24</span></td>
-
 
+
                                          </tr>
-
                            <div class="Title1">Lab Notes</div>
+
                                          <tr>
 +
                                             <td><strong class="style5">Nationality:</strong></td>
 +
                                            <td><span class="style5">Swedish</span></td>
 +
                                          </tr>
 +
                                          <tr>
 +
                                            <td><strong class="style5">Academic background:</strong></td>
 +
                                            <td><span class="style5">B sc. Biotecnology</span></td>
 +
                                          </tr>
 +
                                          <tr>
 +
                                            <td><strong class="style5">Current studies:</strong></td>
 +
                                            <td><span class="style5">Exchange year at University of Minnesota, MS Biotecnology</span></td>
 +
                                          </tr>
 +
                                          <tr>
 +
                                            <td>&nbsp;</td>
 +
                                            <td>&nbsp;</td>
 +
                                          </tr>
 +
                                      </table></td>
 +
                                    </tr>
 +
                                  </table>
 +
      <p>&nbsp;</p>
 +
      <table width="560" border="1">
 +
                                <tr>
 +
                                  <td width="150" rowspan="2" valign="top"><span class="style8"><img src="https://static.igem.org/mediawiki/2010/7/7f/Katarina.jpg" alt="" width="133" height="200" class="img1" /></span></td>  
 +
                                  <td width="151" height="59" valign="top"><table width="150" border="5" cellpadding="0" cellspacing="0">
 +
                                      <tr>
 +
                                        <td><span class="style9">Katarina Risö </span></td>
 +
                                      </tr>
 +
                                      <tr>
 +
                                        <td><span class="style11">student</span></td>
 +
                                      </tr>
 +
                                  </table></td>
 +
                                  <td width="237">&nbsp;</td>
 +
                                </tr>
 +
                                <tr>
 +
                                  <td colspan="2" valign="top"><table width="368" height="150" border="1">
 +
                                    <tr>
 +
                                      <td width="140"><span class="style4">Age:</span></td>
 +
                                      <td width="170"><span class="style5">24</span></td>
 +
                                    </tr>
 +
                                    <tr>
 +
                                      <td><strong class="style5">Nationality:</strong></td>
 +
                                      <td><span class="style5">Swedish</span></td>
 +
                                    </tr>
 +
                                    <tr>
 +
                                      <td><strong class="style5">Academic background:</strong></td>
 +
                                      <td><span class="style5">B sc. Chemical Engineering</span></td>
 +
                                    </tr>
 +
                                    <tr>
 +
                                      <td><strong class="style5">Current studies:</strong></td>
 +
                                      <td><span class="style5">MS Chemistry and Biosciences</span></td>
 +
                                    </tr>
 +
                                    <tr>
 +
                                      <td>&nbsp;</td>
 +
                                      <td>&nbsp;</td>
 +
                                    </tr>
 +
                                  </table>
 +
                                    </td>
 +
                                  </tr>
 +
                              </table>
 +
    <p>&nbsp;</p>
 +
    <table width="560" border="1">
 +
                                  <tr>
 +
                                    <td width="150" rowspan="2" valign="top"><span class="style8"><img src="https://static.igem.org/mediawiki/2010/a/a7/Peidi.jpg" alt="" width="133" height="200" class="img1" /></span></td>
 +
                                    <td width="151" height="59" valign="top"><table width="150" border="5" cellpadding="0" cellspacing="0">
 +
                                        <tr>
 +
                                          <td><span class="style9">Katarina Risö </span></td>
 +
                                        </tr>
 +
                                        <tr>
 +
                                          <td><span class="style11">student</span></td>
 +
                                        </tr>
 +
                                    </table></td>
 +
                                    <td width="237">&nbsp;</td>
 +
                                  </tr>
 +
                                  <tr>
 +
                                    <td colspan="2" valign="top"><table width="368" height="150" border="1">
 +
                                        <tr>
 +
                                          <td width="140"><span class="style4">Age:</span></td>
 +
                                          <td width="170"><span class="style5">24</span></td>
 +
                                        </tr>
 +
                                        <tr>
 +
                                          <td><strong class="style5">Nationality:</strong></td>
 +
                                          <td><span class="style5">Swedish</span></td>
 +
                                        </tr>
 +
                                        <tr>
 +
                                          <td><strong class="style5">Academic background:</strong></td>
 +
                                          <td><span class="style5">B sc. Chemical Engineering</span></td>
 +
                                        </tr>
 +
                                        <tr>
 +
                                          <td><strong class="style5">Current studies:</strong></td>
 +
                                          <td><span class="style5">MS Chemistry and Biosciences</span></td>
 +
                                        </tr>
 +
                                        <tr>
 +
                                          <td>&nbsp;</td>
 +
                                          <td>&nbsp;</td>
 +
                                        </tr>
 +
                                    </table></td>
 +
                                  </tr>
 +
                                </table>
 +
    <p>&nbsp;</p>
 +
    <table width="560" border="1">
 +
                                  <tr>
 +
                                    <td width="150" rowspan="2" valign="top"><span class="style8"><img src="https://static.igem.org/mediawiki/2010/8/80/Adnan.jpg" alt="" width="133" height="200" class="style5" /></span></td>
 +
                                    <td width="151" height="59" valign="top"><table width="150" border="5" cellpadding="0" cellspacing="0">
 +
                                        <tr>
 +
                                          <td><span class="style9">Adnan Kadic</span></td>
 +
                                        </tr>
 +
                                        <tr>
 +
                                          <td><span class="style11">student</span></td>
 +
                                        </tr>
 +
                                    </table></td>
 +
                                    <td width="237">&nbsp;</td>
 +
                                  </tr>
 +
                                  <tr>
 +
                                    <td colspan="2" valign="top"><table width="368" height="150" border="1">
 +
                                        <tr>
 +
                                          <td width="140"><span class="style4">Age:</span></td>
 +
                                          <td width="170"><span class="style5">24</span></td>
 +
                                        </tr>
 +
                                        <tr>
 +
                                          <td><strong class="style5">Nationality:</strong></td>
 +
                                          <td><span class="style5">Bosnian</span></td>
 +
                                        </tr>
 +
                                        <tr>
 +
                                          <td><strong class="style5">Academic background:</strong></td>
 +
                                          <td><span class="style5">B sc. Biotecnology</span></td>
 +
                                        </tr>
 +
                                        <tr>
 +
                                          <td><strong class="style5">Current studies:</strong></td>
 +
                                          <td><span class="style5">MS Chemistry and Biosciences </span></td>
 +
                                        </tr>
 +
                                        <tr>
 +
                                          <td>&nbsp;</td>
 +
                                          <td>&nbsp;</td>
 +
                                        </tr>
 +
                                    </table></td>
 +
                                  </tr>
 +
                                </table>
 +
    <p>&nbsp;</p>
 +
    <table width="560" border="1">
 +
                                  <tr>
 +
                                    <td width="150" rowspan="2" valign="top"><span class="style8"><img src="https://static.igem.org/mediawiki/2010/e/ea/Lokesh.jpg" alt="" width="133" height="200" class="style5" /></span></td>
 +
                                    <td width="221" height="59" valign="top"><table width="216" border="5" cellpadding="0" cellspacing="0">
 +
                                        <tr>
 +
                                          <td width="180"><span class="style9">Lokeshwaran Manoharan </span></td>
 +
                                        </tr>
 +
                                        <tr>
 +
                                          <td><span class="style11">student</span></td>
 +
                                        </tr>
 +
                                    </table></td>
 +
                                    <td width="124">&nbsp;</td>
 +
                                  </tr>
 +
                                  <tr>
 +
                                    <td colspan="2" valign="top"><table width="368" height="150" border="1">
 +
                                        <tr>
 +
                                          <td width="140"><span class="style4">Age:</span></td>
 +
                                          <td width="170"><span class="style5">23</span></td>
 +
                                        </tr>
 +
                                        <tr>
 +
                                          <td><strong class="style5">Nationality:</strong></td>
 +
                                          <td><span class="style5">Indian</span></td>
 +
                                        </tr>
 +
                                        <tr>
 +
                                          <td><strong class="style5">Academic background:</strong></td>
 +
                                          <td><span class="style5">B.Tech Ind Biotechnology</span></td>
 +
                                        </tr>
 +
                                        <tr>
 +
                                          <td><strong class="style5">Current studies:</strong></td>
 +
                                          <td><span class="style5">MS Bioinformatics</span></td>
 +
                                        </tr>
 +
                                        <tr>
 +
                                          <td>&nbsp;</td>
 +
                                          <td>&nbsp;</td>
 +
                                        </tr>
 +
                                    </table></td>
 +
                                  </tr>
 +
                                </table>
 +
    <p>&nbsp;</p>
 +
    <table width="560" border="1">
 +
                                  <tr>
 +
                                    <td width="150" rowspan="2" valign="top"><span class="style8"><img src="https://static.igem.org/mediawiki/2010/0/01/Kemal.jpg" alt="" width="133" height="200" class="style5" /></span></td>
 +
                                    <td width="207" height="59" valign="top"><table width="186" border="5" cellpadding="0" cellspacing="0">
 +
                                         <tr>
 +
                                          <td width="150"><span class="style9">Kemal Sanli </span></td>
 +
                                        </tr>
 +
                                        <tr>
 +
                                          <td><span class="style11">student</span></td>
 +
                                        </tr>
 +
                                     </table></td>
 +
                                    <td width="161">&nbsp;</td>
 +
                                  </tr>
 +
                                  <tr>
 +
                                    <td colspan="2" valign="top"><table width="368" height="150" border="1">
 +
                                        <tr>
 +
                                          <td width="133"><span class="style4">Age:</span></td>
 +
                                          <td width="177"><span class="style5">24</span></td>
 +
                                        </tr>
 +
                                        <tr>
 +
                                          <td><strong class="style5">Nationality:</strong></td>
 +
                                          <td><span class="style5">Turkish</span></td>
 +
                                        </tr>
 +
                                        <tr>
 +
                                          <td><strong class="style5">Academic background:</strong></td>
 +
                                          <td><span class="style5">B sc. Molecular Biology and Genetics </span></td>
 +
                                        </tr>
 +
                                        <tr>
 +
                                          <td><strong class="style5">Current studies:</strong></td>
 +
                                          <td><span class="style5">MS Bioinformatics</span></td>
 +
                                        </tr>
 +
                                        <tr>
 +
                                          <td>&nbsp;</td>
 +
                                          <td>&nbsp;</td>
 +
                                        </tr>
 +
                                    </table></td>
 +
                                  </tr>
 +
                                </table>
 +
    <p>&nbsp;</p>
 +
    <table width="560" border="1">
 +
                                  <tr>
 +
                                    <td width="150" rowspan="2" valign="top"><span class="style8"><img src="https://static.igem.org/mediawiki/2010/4/40/Karl.jpg" alt="" width="133" height="200" class="style5" /></span></td>
 +
                                    <td width="207" height="59" valign="top"><table width="186" border="5" cellpadding="0" cellspacing="0">
 +
                                        <tr>
 +
                                          <td width="150"><span class="style9">Karl Almén Burman </span></td>
 +
                                        </tr>
 +
                                        <tr>
 +
                                          <td><span class="style11">student</span></td>
 +
                                        </tr>
 +
                                    </table></td>
 +
                                    <td width="161">&nbsp;</td>
 +
                                  </tr>
 +
                                  <tr>
 +
                                    <td colspan="2" valign="top"><table width="368" height="150" border="1">
 +
                                        <tr>
 +
                                          <td width="133"><span class="style4">Age:</span></td>
 +
                                          <td width="177"><span class="style5">23</span></td>
 +
                                        </tr>
 +
                                        <tr>
 +
                                          <td><strong class="style5">Nationality:</strong></td>
 +
                                          <td><span class="style5">Swedish</span></td>
 +
                                        </tr>
 +
                                        <tr>
 +
                                          <td><strong class="style5">Academic background:</strong></td>
 +
                                          <td><span class="style5">B sc. Biotechnology </span></td>
 +
                                        </tr>
 +
                                        <tr>
 +
                                          <td><strong class="style5">Current studies:</strong></td>
 +
                                          <td><span class="style5">MS Biotechnology</span></td>
 +
                                        </tr>
 +
                                        <tr>
 +
                                          <td>&nbsp;</td>
 +
                                          <td>&nbsp;</td>
 +
                                        </tr>
 +
                                    </table></td>
 +
                                  </tr>
 +
                                </table>
 +
    <p>&nbsp;</p>
 +
              <table width="560" border="1">
 +
                                    <tr>
 +
                                      <td width="150" rowspan="2" valign="top"><span class="style8"><img src="https://static.igem.org/mediawiki/2010/e/e8/Julia.jpg" alt="" width="133" height="200" class="style5" /></span></td>
 +
                                      <td width="207" height="59" valign="top"><table width="186" border="5" cellpadding="0" cellspacing="0">
 +
                                          <tr>
 +
                                            <td width="150"><span class="style9">Julia Fransson </span></td>
 +
                                          </tr>
 +
                                          <tr>
 +
                                            <td><span class="style11">student</span></td>
 +
                                          </tr>
 +
                                      </table></td>
 +
                                      <td width="161">&nbsp;</td>
 +
                                    </tr>
 +
                                    <tr>
 +
                                      <td colspan="2" valign="top"><table width="368" height="150" border="1">
 +
                                          <tr>
 +
                                            <td width="133"><span class="style4">Age:</span></td>
 +
                                            <td width="177"><span class="style5">22</span></td>
 +
                                          </tr>
 +
                                          <tr>
 +
                                            <td><strong class="style5">Nationality:</strong></td>
 +
                                            <td><span class="style5">Swedish</span></td>
 +
                                          </tr>
 +
                                          <tr>
 +
                                            <td><strong class="style5">Academic background:</strong></td>
 +
                                            <td><span class="style5">B sc. Biotechnology </span></td>
 +
                                          </tr>
 +
                                          <tr>
 +
                                            <td><strong class="style5">Current studies:</strong></td>
 +
                                            <td><span class="style5">Exchange year through Unitech International</span></td>
 +
                                          </tr>
 +
                                          <tr>
 +
                                            <td>&nbsp;</td>
 +
                                            <td>&nbsp;</td>
 +
                                          </tr>
 +
                                      </table></td>
 +
                                    </tr>
 +
                                  </table>
 +
              <p>&nbsp;</p>
 +
              </td>
 +
                                  </tr>
 +
                                  <tr><td>&nbsp;</td>
 +
  </tr><tr><td>&nbsp;</td>
 +
  </tr>
 +
    </div>
 +
 +
</div>
 +
</div>
 +
<div id="right">
 +
                <div class="Title3">Lab Notes</div>
                            
                            
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                              <div class="left_grad">
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                                    <div class="lab">
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                                      <div id="lab">
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                                        <div class="numleft">
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                                        <div class="numleft">
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                                          <p>11</p>
+
                                          <p>11</p>
                                         </div>
                                         </div>
-
                                        <div class="newsright"><a href="#">August 11, 2010</a>
+
                                          <div class="newsright"><a href="#">August 11, 2010</a>
-
                                            <p2>The PCR reaction from yesterday was checked on a gel. α should be ... </p2>
+
                                          <p> The PCR reaction from yesterday was checked on a gel. α should be ... </p>
-
                                        </div>
+
                                          </div>
-
                                        <div class="read"><a href="#">read more</a></div>
+
 
-
                                        <div style="clear: both"></div>
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                                          <div class="read"><a href="#">read more</a></div>
-
                                     </div>
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                                          <div style="clear: both"></div>
-
                                    <div>
+
                                      
-
                                        <div class="numleft"><p>10</p></div>
+
           
-
                                        <div class="newsright"><a href="#">August 10, 2010</a>
+
                               
-
                                            <p2>The transformation plates were checked but nothing had grown. Prepared PCR reaction Γ ...  </p2>
+
    </div>    
-
                                        </div>
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-
                                     
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</div>
-
                               
+
-
                                    </div>  <div class="read"><a href="#">read more</a></div>
+
  <div id="myfooter"> <p>   <a href="#">Home</a> | <a href="#">CTH</a> | <a href="#">Sponsors</a> | <a href="#">About Us</a> | <a href="#">Contact Us</a></p> <p>Chalmers University of Technology, Gothenburg, SWEDEN</p>
-
<div class="list_border"></div>  
+
-
                              </div>
+
              </div>
-
                            </div>
+
</div></div>  
-
                        </div>
+
</body>
-
+
-
                        <div id="center">
+
-
                            <div id="Title2">Preliminary</div>
+
-
                            <div class="text">
+
-
                             
+
-
                              <strong><p>Fluorescent  Proteins:</p></strong>
+
-
    <p>Green Fluorescent Protein <a href="https://2010.igem.org/Team:Gothenburg-Sweden/Lab_Note/pre#FP">(read more)</a></p>
+
-
<strong><p>Plasmid backbone:</p></strong>
+
-
                    <p>pSP-GM1 <a href="https://2010.igem.org/Team:Gothenburg-Sweden/Lab_Note/pre#plasmid">(read more)</a></p>
+
-
    <strong><p>SNF1  (modified subunits):</p></strong>
+
-
                    <p>Snf1 (alfa), Snf4 (gamma) <a href="https://2010.igem.org/Team:Gothenburg-Sweden/Lab_Note/pre#SNF1">(read more)</a></p>
+
-
    <strong><p>FP  positions:</p></strong>
+
-
                    <p>N-terminal alfa with N-terminal gamma, N-terminal gamma with C-terminal gamma <a href="https://2010.igem.org/Team:Gothenburg-Sweden/Lab_Note/pre#position">(read more)</a></p>
+
-
    <strong><p>Primer  design:</p></strong>
+
-
                    <p>Fusion primers with restiriction enzyme sites included at very ends, each annealing part was desinged with an melting temperature of around 60 C <a href="https://2010.igem.org/Team:Gothenburg-Sweden/Lab_Note/pre#primer">(read more)</a></p>
+
-
 
+
-
                          </div>
+
-
                            <div id="Title2">Lab work by date</div>
+
-
                           
+
-
                            <div class="text">    <ul><li><p> <strong>2010-08-11</strong><br>
+
-
                              Malin and Katarina<br>
+
-
                              <br>
+
-
                              The PCR reaction from yesterday was checked on a gel. α should be around 1500 bp but only a band around 750 bp was seen. We had a long meeting with supervisor Per about PCR reactions and the next couple of weeks in the lab. <br>
+
-
                              <br>
+
-
                              Karl and Kemal <br>
+
-
                              <br>
+
-
                              Ran PCR reaction α with different settings. <br>
+
-
                              <strong>α1: </strong>regular settings<br>
+
-
                              <strong>α2: </strong>10 x diluted template 2<br>
+
-
                              <strong>α3: </strong>increased annealing time from 30 s to 60 s<br>
+
-
                              <strong>α4: </strong>decreased annealing temperature<br>
+
-
                              <br>
+
-
                              After PCR, the products were run on a gel. α1, α3 and α 4 had a band around 750 bp but nothing on α2. </p></li><br>
+
-
                              <br>
+
-
                              <li><p> <strong>2010-08-10</strong><br>
+
-
                              Peidi and Lokesh<br>
+
-
                              <br>
+
-
                              Checked the transformation plates but nothing had grown. Prepared PCR reaction Γ. <br>
+
-
                              <br>
+
-
                              Katarina and Kemal<br>
+
-
                              <br>
+
-
                              PCR product Γ was verified on a 0.7 % agarose gel. There was a band around 750 bp as expected and it was purified using the regular protocol. A new phusion mastermix was made and PCR reaction α was run with Γ as one of the template. Regular phusion PCR settings was used. <br>
+
-
                              <br></p></li>
+
-
                              <li><p><strong>2010-08-09</strong><br>
+
-
                              Malin and Kemal<br>
+
-
                              <br>
+
-
                              Before checking if the ligation worked we need to transform and amplify in E.coli. Then we can miniprep the ligated plasmid and check on a gel. After that we can digest plasmid with PacI and BcuI and ligate the digested IV. Then another round of transformation and miniprep before we can transform yeast. <br>
+
-
                              <br>
+
-
                              Karl and Lokesh<br>
+
-
                              <br>
+
-
                              Transformed competent E.coli with plasmid ligated with III by heat shock. The cells were incubated at 37 °C over night. <br>
+
-
                              <br></p></li>
+
-
                              <li><p> <strong>2010-08-07</strong><br>
+
-
                              Lokesh<br>
+
-
                              <br>
+
-
                              Moved ligation tube to freezer. <br>
+
-
                              <br></p></li>
+
-
                            <li><p>  <strong>2010-08-06</strong><br>
+
-
                              Malin, Julia and Lokesh<br>
+
-
                              <br>
+
-
                              III and IV were digested 3x protocol and were then put on a 1 % agarose gel together with Γ both for verification and purification. Correct bands were seen for purification except for Γ which was empty. In the verification lanes the digested PCR products were to light to be seen but they were seen in purification lanes. IIId and IVd were purified. <br>
+
-
                              <br>
+
-
                              <strong>IIId: </strong>7.8 ng/µl<br>
+
-
                              <strong>IVd: </strong>11.7ng/µl<br>
+
-
                              <br>
+
-
                              IIId will now be ligated with the cut plasmid (BamHI and HindIII). Ligation performed according to protocol with 156 ng insert and 52 ng plasmid. Ligation was incubated over night in fridge at 4-16° C. <br>
+
-
                              <br>
+
-
                              All 4 sporulation plates were checked but still no tetrads visible. <br>
+
-
                              <br></p></li>
+
-
                              <li><p>  <strong>2010-08-05</strong><br>
+
-
                              Malin and Julia<br>
+
-
                              <br>
+
-
                              III and IV were digested using the regular protocol. <br>
+
-
                              <strong>III: </strong>HindIII and BamHI works with Buffer B<br>
+
-
                              <strong>IV: </strong>PacI and BcuI works with Buffer M<br>
+
-
                              <br>
+
-
                              A 1% gel was run with uncut and digested III and IV, control, I and Β. I worked well and will be purified. The Β lane showed no bands. None of the digested products were visible on the gel so a new gel was run trying to verify this result. The control shows a band around 1700 which is weird. This band is also visible on the III uncut but together with a band around 750 bp. No bands at the IV which is very strange since this PCR product has been verified before. Have something gone wrong with the purification? <br>
+
-
                              <br>
+
-
                              As a result PCR reactions III and IV will be redone. New PCR reactions of Γ and α were also performed in the same block. <br>
+
-
                              <br>
+
-
                              Karl and Kemal<br>
+
-
                              <br>
+
-
                              Reactions of Β were made. Β1 is a continuation of the 100 cycle PCR this time with primers. Β2 uses a megaprimer approach. <br>
+
-
                              <br>
+
-
                              All the PCR products were run on a 1 % agarose gel. III, IV and Γ were alright. α and Β2 showed a band around 750 bp. <br>
+
-
                              <strong>III: </strong>531.3 ng/µl<br>
+
-
                              <strong>IV: </strong>263.9 ng/µl<br>
+
-
                              <br></p></li>
+
-
                              <li><p> <strong>2010-08-04</strong><br>
+
-
                              Julia and Karl<br>
+
-
                              <br>
+
-
                              PCR products Β were loaded on a 0.7 % gel. No correct bands, only amplified templates around 750 bp and something else around 500 bp. <br>
+
-
                              <br>
+
-
                              PCR of SAMS will also be redone from scratch. PCR reactions 1, 2 and C are done, then loaded on a gel (2 % gel and low range ladder) and purified. <br>
+
-
                              <strong>1: </strong>8.2 ng/µl<br>
+
-
                              <strong>2: </strong>7.8 ng/µl<br>
+
-
                              <strong>C: </strong>4.9 ng/µl<br>
+
-
                              Samples diluted to 1 ng/µl. <br>
+
-
                              <br>
+
-
                              PCR of Β was done again but this time in two steps and with the Finnzymes Tm which corresponds to an annealing temp at 85 °C. Firstly 10 cycles without primers were performed. <br>
+
-
                              <br>
+
-
                              Kemal and Lokesh<br>
+
-
                              <br>
+
-
                              Second round of PCR with primers were performed. Then new PCR reactions amplifying III and IV were done and also reaction I. <br>
+
-
                              <strong>III: </strong>572 ng/µl<br>
+
-
                              <strong>IV: </strong>660 ng/µl<br>
+
-
                              <br>
+
-
                              The sporplates were checked and very few tetrads were seen. <br>
+
-
                              <br>
+
-
                              Β samples were put on a gel but no correct bands visible. We suspect that the reverse primer is attaching in the wrong place since YFP and CFP is so similar which then would lead to 750 bp fragment. <br>
+
-
                              <br></p></li>
+
-
                              <li><p>  <strong>2010-08-03</strong><br>
+
-
                              Malin and Lokesh<br>
+
-
                              <br>
+
-
                              PCR products II, III and IV were verified and purified from a 1 % agarose gel. III did not work and was not purified. <br>
+
-
                              <br>
+
-
                              Concentrations: <br>
+
-
                              <strong>II: </strong>6.2 ng/µl <br>
+
-
                              <strong>IV: </strong>5.6 ng/µl<br>
+
-
                              II and IV were diluted to 1 ng/µl. <br>
+
-
                              <br>
+
-
                              The new sporplates were examined but no tetrads visible. To check further the cells were stained with DAPI and examined in fluorescence microscope. Still no success. <br>
+
-
                              <br>
+
-
                              New PCR reactions were prepared. III was redone and Β was also done. Same settings as yesterday. <br>
+
-
                              <br>
+
-
                              Kemal and Karl<br>
+
-
                              <br>
+
-
                              PCR products were loaded on a 1% agarose gel for verification and purification. III worked this time and was purified from the gel using the protocol from QIAquick Gel Extraction Kit. <br>
+
-
                              <strong>III: </strong>6.4 ng/µl<br>
+
-
                              <br>
+
-
                              New PCR of Β was run with two different block settings. <br>
+
-
                              <strong>Block A: </strong>increased annealing time to 1 min. <br>
+
-
                              <strong>Block B: </strong>increased annealing time to 58 °C. <br>
+
-
                              <br></p></li>
+
-
                              <li><p> <strong>2010-08-02</strong><br>
+
-
                              Kemal, Lokesh and Karl<br>
+
-
                              <br>
+
-
                              PCR products A, B and 3 were loaded on a gel and then purified from the gel using the protocol from QIAquick Gel Extraction Kit. α and IV were also run on a gel but only showed strong bands corresponding to amplified templates. On the α lane however a very light band was seen at the correct length. This will be further investigated. <br>
+
-
                              <br>
+
-
                              Malin and Julia<br>
+
-
                              <br>
+
-
                              The purified PCR products were measured with nanodrop: <br>
+
-
                              <strong>A1: </strong>11.8 ng/µl<br>
+
-
                              <strong>A2: </strong>10.2 ng/µl<br>
+
-
                              <strong>B1: </strong>9.6 ng/µl<br>
+
-
                              <strong>B2: </strong>17.1 ng/µl<br>
+
-
                              <strong>31: </strong>15.0 ng/µl<br>
+
-
                              <strong>32: </strong>16.3 ng/µl<br>
+
-
                              <br>
+
-
                              A, B and 3 were diluted to 1 ng/µl. <br>
+
-
                              <br>
+
-
                              PCR products 4 and 5 were loaded on a 1 % agarose gel together with IV that should be controlled. PCR products 4 and 5 were correct so they were purified from the gel using the protocol from QIAquick Gel Extraction Kit. IV however was not correct. Only a band around 750 bp was visible. <br>
+
-
                              <br>
+
-
                              Concentrations: <br>
+
-
                              <strong>4: </strong>10.8 ng/µl<br>
+
-
                              <strong>5: </strong>10.4 ng/µl<br>
+
-
                              4-5 were diluted to 1 ng/µl. <br>
+
-
                              New PCR reactions were started (II, III and IV). PCR settings that works well with the Phusion enzyme was set: <br>
+
-
                              Initial denaturation: 98 °C 3 min<br>
+
-
                              Denaturation: 98 °C 10 s<br>
+
-
                              Annealing: 55 °C 30 s<br>
+
-
                              Elongation: 72 °C 1.5 min+ 1s/cycle<br>
+
-
                              30 cycles<br>
+
-
                              Final extension: 72 °C 10 min<br>
+
-
                              4°C forever <br>
+
-
                              <br>
+
-
                              <strong>2010-07-30</strong><br>
+
-
                              Julia and Katarina<br>
+
-
                              <br>
+
-
                              The concentration of the purified plasmids were measured with nanodrop: <br>
+
-
                              <strong>P1: </strong>1.7 ng/µl<br>
+
-
                              <strong>P2: </strong>3.5 ng/µl<br>
+
-
                              <strong>P3: </strong>25.9 ng/µl<br>
+
-
                              <strong>P4: </strong>42.8 ng/µl<br>
+
-
                              This is clear evidence that water should be used for eluation in the future! <br>
+
-
                              <br>
+
-
                              To make sure that we have the right products after purification a 1 % agarose gel was run with fastruler DNA ladder. The gel verified the plasmid size though the bands were light. <br>
+
-
                              <br>
+
-
                              PCR product III was evaporated since it was so dilute. New concentration: <br>
+
-
                              <strong>IIId: </strong>35 ng/µL<br>
+
-
                              <br>
+
-
                              The digested PCR product III and the digested plasmid were ligated with T4 ligase. The DNA concentration in total should not exceed 1 µg. The ratio insert:vector should be 3:1. <br>
+
-
                              <br>
+
-
                              <strong>Ligation protocol: </strong><br>
+
-
                              150 ng insert<br>
+
-
                              50 ng vector<br>
+
-
                              3 µl 10x T4 buffer<br>
+
-
                              2.5 µl (2.5 U, between 1-5 U) T4 ligase<br>
+
-
                              Adjust volume with water up to 30 µl. Mix and incubate in 4-16 °C (refrigerator) overnight. <br>
+
-
                              The newly plated sporplates were moved to 30°C. The old sporplates were again studied in the microscope and no tetrads were seen. <br>
+
-
                              <br>
+
-
                              Kemal and Lokesh<br>
+
-
                              <br>
+
-
                              Since we have had a lot of trouble with the longer fusions we will start over from scratch with a new high fidelity polymerase called “Finnzumes Phusion”. New PCR reactions were run (A, B, 3, 4 and 5). IV and α were also performed using alternative settings (2 different PCR reactions described before) but with “old” templates. <br>
+
-
                              <br></p></li>
+
-
                              <li><p><strong>2010-07-29</strong><br>
+
-
                              Julia and Katarina<br>
+
-
                              <br>
+
-
                              PCR product III is the final fusion protein where Snf4 is tagged with yfp. To be able to insert the construct into the plasmid we must digest it. <br>
+
-
                              <br>
+
-
                              Digestion: Mix DNA, buffer and water. Last add enzyme and mix. Incubate at 37° C for 1 hour. <br>
+
-
                              1 µg DNA<br>
+
-
                              2.5 µl Buffer<br>
+
-
                              1 U Enzyme<br>
+
-
                              Up to 25 µl H2O<br>
+
-
                              For PCR product III, the restriction enzymes BamHI and HindIII are used together with buffer B. After digestion (the protocol was scaled 3x) the sample was loaded on a 1 % agarose gel in 6 lanes with 1 kb ladder. The bands were cut out and weighed. The samples were purified from the gel using the protocol from QIAquick Gel Extraction Kit. The DNA was eluated with water. <br>
+
-
                              <br>
+
-
                              Concentration: <br>
+
-
                              <strong>IIId: </strong>5 ng/µl (Total product 160 µl= 800 ng) <br>
+
-
                              <br>
+
-
                              Kemal and Lokesh<br>
+
-
                              <br>
+
-
                              The plasmids were also digested with BamHI and HindIII according to the same protocol as above. P1 and P2 were eluated with buffer EB and P3 and P4 with water. <br>
+
-
                              <br>
+
-
                              2 new sporulation plates were prepared by plating the diploid yeast cells on spore plates and kept at room temperature. <br>
+
-
                              <br></p></li>
+
-
                                <li><p><strong>2010-07-28</strong><br>
+
-
                              Julia and Katarina<br>
+
-
                              <br>
+
-
                              The PCR products were run on a 1 % gel with 1 kb ladder. Γ may have worked. α and Β had a band around 750 bp and a weird light band around 500 bp. They obviously did not work. IV did not work at all. <br>
+
-
                              <br>
+
-
                              The sporulation plates were checked for spores. Cells were studied under a microscope. No tetrads visible. <br>
+
-
                              <br></p></li>
+
-
                              <li><p> <strong>2010-07-27</strong><br>
+
-
                              Julia and Katarina<br>
+
-
                              <br>
+
-
                              A 1.5 % gel was run with all of the fragments around 700 bp to control the validity of the PCR reactions. A low range ladder was used. PCR products 1-5 seems good. The results from yesterday were confirmed. <br>
+
-
                              <br>
+
-
                              A new PCR design was made with α, Β, Γ and IV. PCR was done in two steps to increase the chance of success for longer fusions. <br>
+
-
                              1. 7 cycles with short annealing time. <br>
+
-
                              2. 20 cycles with long annealing time (all other settings kept standard). <br>
+
-
                              <br></p></li>
+
-
                                <li><p><strong>2010-07-26</strong><br>
+
-
                              <br>
+
-
                              A new experiment were designed called Γ which is the same as I but with longer annealing region. Also an experiment called Δ were designed which corresponds to II. <br>
+
-
                              <br>
+
-
                              Julia and Katarina<br>
+
-
                              <br>
+
-
                              PCR reactions were run with samples of 4, IV and Γ. IV was tried with two different PCR settings: one regular and one with two runs of PCR, the first without primers and only 5 cycles and the second one with regular settings. <br>
+
-
                              <br>
+
-
                              Kemal and Lokesh<br>
+
-
                              <br>
+
-
                              The PCR products were controlled on a 0.7% agarose gel. Only 4 worked. Γ was empty and IV had bands in the wrong place (amplified template). <br>
+
-
                              <strong>4: </strong>534 ng/ µl </p></li>
+
-
                                <li><p>  <strong>2010-07-22</strong><br>
+
-
                                      Karl, Lokesh and Kemal
+
-
                                      </br></br>
+
-
                                      A new mastermix was prepared using a different enzyme. Then the  experiments from yesterday was redone using regular PCR settings: <br>
+
-
                                      A, 5, 2, I<br>
+
-
                                      Then some more experiments of IV was redone: <br>
+
-
                                      2a, 2b and 2c<br>
+
-
                                      The new primer 14 had arrived so PCR reaction 4 was also redone. <br>
+
-
                                      <br>
+
-
                                      All the PCR products were run on a 0.7 % agarose gel with GeneRuler 1 kb  DNA ladder. The results were inconclusive. None of the expected bands  were received.  At this point we are not sure what to do since none of  the PCR reactions worked. <br>
+
-
                                      <br>
+
-
                                      </p>
+
-
                                </li>
+
-
                                <li> <p> <strong>2010-07-21</strong><br>
+
-
                                  Malin, Julia, Karl and Lokesh<br>
+
-
                                  <br>
+
-
                                  <strong>Verification of the PCR products </strong>α1, α2, α3, α4, α5 and αK by running on a thin 0.7 % agarose gel (8 µl sample+ 2 µl loading buffer, GeneRuler 1 kb DNA ladder). <br>
+
-
                                  <br>
+
-
                                  The gel didn’t show the expected band lengths. A band around 700 bp  could be seen, most strongly for α4 and α5.  We suspect that there is  something wrong with the ladder because we get inconclusive results.  Since none of the PCR reactions for the last couple of days have worked  we will redo the templates of α and IV: I, 2, 5 and A. These PCR  reactions have worked before. <br>
+
-
                                  <br>
+
-
                                  <strong>Expected fragment length: </strong><br>
+
-
                                  A.2: 1938 bp<br>
+
-
                                  5.2: 757 bp<br>
+
-
                                  2.2: 762 bp<br>
+
-
                                  I.2: 768 bp<br>
+
-
                                  <br>
+
-
                                  The PCR products were run on a thin 0.7 % agarose gel. We used both the  FastRuler High Range Ladder and the GeneRuler 1 kb DNA ladder. 5.2 and  2.2 showed the expected bands around 750 bp. I.2 and A.2 did not work at  all, nothing visible on the gel.  We also run some of the old PCR  products to see what the result would be using a new ladder. We run α2,  α5, 4c (IV) and 4c*(IV). Here some template amplifications were visible  but sadly none of the expected fragment lengths. Both ladders showed the  same result so there is probably nothing wrong with the GeneRuler  ladder except that it is not showing strong color. <br></p></li>
+
-
                                  <li><p><strong>2010-07-20</strong><br>
+
-
                                  Malin, Julia, Lokesh and Karl<br>
+
-
                                  <br>
+
-
                                  <strong>Verification of yesterdays PCR reactions </strong>by running on a 0.7 % agarose gel (8 µl sample+ 2 µl loading buffer, GeneRuler 1 kb DNA ladder) <br>
+
-
                                  <br>
+
-
                                  None of the reactions worked. <br>
+
-
                                  <br>
+
-
                                  Gel 1 (Block A): Nothing visible on gel, ladders only very light so  these samples were run on a new 0.7 % agarose gel but this time thinner  to get better resulotion (0.58 g agarose + 80 ml TBE). This time no  expected fragments were seen, but bands corresponding to primer dimers  were clearly seen. <br>
+
-
                                  Gel 2 (Block B): A few small bands were seen but no bands of expected length. <br>
+
-
                                  <br>
+
-
                                  We decided to stop working on IV until proper feedback on the experimental setup was received. Instead we continued with α.<br>
+
-
                                  <br>
+
-
                                  <strong>Doing another PCR round of α (SAMS peptide): </strong> <br>
+
-
                                  <strong>α: PCR fusion of cfp-SAMS and yfp: </strong>Adds BamHI on N-terminus of cfp and HindIII on C-terminus of yfp and keeps the stop codons. Expected fragment length: 1502 bp. <br>
+
-
                                  <br>
+
-
                                  α1: regular protocol. <br>
+
-
                                  α2: regular protocol. <br>
+
-
                                  α3: 1.25 µl extra primer 9.<br>
+
-
                                  α4: 1.25 µl extra primer 12. <br>
+
-
                                  α5: 1.25 µl extra primer 9 and 1.25 µl primer 12. <br>
+
-
                                  αK: Control<br>
+
-
                                  Regular PCR-settings</p></li>
+
-
                                <li><p> <strong>2010-07-19</strong><br>
+
-
                                  Karl, Peidi and Kemal<br>
+
-
                                  <br>
+
-
                                  <strong>Doing a new experimental setup for IV: </strong><br>
+
-
                                  <strong>IV. PCR fusion of cfp and Snf1: </strong>Adds SpeI on N-terminus of cfp and PacI on C-terminus of Snf1 and keeps the stop codon. Expected fragment length: 2643 bp. <br>
+
-
                                  <br>
+
-
                                  13 different tubes were prepared and run at two different PCR-settings.  Block A: Regular settings except double annealing time (1 min). Block B:    Regular settings except 58° C annealing temperature. <br>
+
-
                                  <br>
+
-
                                  C : Control<br>
+
-
                                  1a: same as 2* (template 5 5 x diluted) <br>
+
-
                                  1b. same as 3* (template 5 10 x diluted) <br>
+
-
                                  1c: same as 4* (template 5 100x diluted) <br>
+
-
                                  2a: increased product A concentration to 5 ng/µl. <br>
+
-
                                  2b: increased product A concentration to 10 ng/µl. <br>
+
-
                                  3a: same as 5* (2x primer 7) <br>
+
-
                                  3b: same as 6* (template 5 diluted 5x, 2x primer 7) <br>
+
-
                                  3c: same as 7* (template 5 diluted 10x, 2x primer 7) <br>
+
-
                                  3d same as 8* (template 5 diluted 100x, 2x primer 7) <br>
+
-
                                  4a: 2x primer 7. <br>
+
-
                                  4b: 2x primer 7, 5x template A. <br>
+
-
                                  4c: 2x primer 7, 10 x template A. <br></p></li>
+
-
                                <li><p> <strong>2010-07-16</strong><br>
+
-
                                  Lokesh, Kemal, Katarina and Peidi<br>
+
-
                                  <br>
+
-
                                  <strong>Verification of PCR </strong>reactions IV* (8 tubes) and α on a 0.7 % agarose gel (5 µl sample+ 2µl loading buffer, GeneRuler 1 kb DNA ladder). <br>
+
-
                                  <br>
+
-
                                  No bands visible. None of the PCR reactions worked, just small bands  visible corresponding to primers. The α PCR reaction messed the tubes  up. That might be an explanation why there were no bands on the gel for  α.  A new experimental setup must be made tomorrow. <br>
+
-
                                  <br>
+
-
                                  PCR products I and 2 were run with a low range ladder to see that those products are ok. That gel was satisfactory. <br>
+
-
                                  <br></p>
+
-
                                  <li><p><strong>2010-07-15</strong><br>
+
-
                                  Malin, Julia, Kemal and Karl<br>
+
-
                                  <br>
+
-
                                  <strong>Concentrations of PCR products: </strong><br>
+
-
                                  4.2. 356.2 ng/µl<br>
+
-
                                  I. 338.3 ng/µl<br>
+
-
                                  II. 303.6 ng/µl<br>
+
-
                                  III. 337.5 ng/µl<br>
+
-
                                  IV. 280.9 ng/µl<br>
+
-
                                  <br>
+
-
                                  All PCR-products were diluted to 1 ng/µl. <br>
+
-
                                  <br>
+
-
                                  <strong>Verified PCR products</strong> I-IV and 4.2 on a 0.7 % Agarose Gel (8 µl sample + 2µl loading buffer, GeneRuler 1 kb DNA ladder) <br>
+
-
                                  PCR products I, II and III were good but 4.2 showed no bands at all.  After primer design control we found a problem with primer 14, which  then was ordered again. IV had only a band around 700 bp instead of  2600.  This might be template 5, which then has been amplified. If the  settings are changed maybe it will work. Firstly the template 5 will be  diluted 0x, 5x, 10x and 100x (4 tubes: 1*, 2*, 3* and 4*) and these  dilutions will also be run with higher primer concentration 7 (4 tubes:  5*, 6*, 7* and 8*).  2.5 µl primer 7 will be added instead of 1.25 µl  primer 7. <br>
+
-
                                  <br>
+
-
                                  New PCR reactions were run: <br>
+
-
                                  <strong>α: PCR fusion of cfp-SAMS and yfp: </strong> Adds BamHI on N-terminus of cfp and HindIII on C-terminus of yfp and keeps the stop codons. Expected fragment length: 1502 bp. <br>
+
-
                                  K4. Control. <br>
+
-
                                  <br>
+
-
                                  <strong>IV *: PCR fusion of cfp and Snf1: </strong>Adds SpeI on N-terminus of cfp and PacI on C-terminus of Snf1 and keeps the stop codon. Expected fragment length: 2643 bp. <br>
+
-
                                  Half volume 25 µl. Regular PCR settings except 56 °C annealing temperature. <br>
+
-
                                  1*;- 8*<br>
+
-
                                  K1*. Control for 1-4*.<br>
+
-
                                  K2*. Control for 5-8*.<br>
+
-
                                  <br></p></li>
+
-
                             
+
-
                                <li> <p><strong>2010-07-14</strong><br>
+
-
                                  Julia and Katarina <br>
+
-
                                  <strong>Ran PCR 1-5.</strong><br>
+
-
                                  1. Amplification of cfp-sams compatible: Add restriction enzyme site BamHI on N-terminus +SAMS on C-terminus.<br>
+
-
                                  Expected fragment length: 748 bp.<br>
+
-
                                  2. Amplification of yfp-SAMS competible: Adds restriction enzyme site for HindIII on C-terminus and SAMS tail on N-terminus. <br>
+
-
                                  Expected fragment length: 762 bp.<br>
+
-
                                  3. Amplification of yfp-snf4 compatible: Adds restriction enzyme site BamHI on N-terminus and Snf4 tail on C-terminus. <br>
+
-
                                  Expected fragment length: 755 bp.<br>
+
-
                                  4. Amplification of cfp-Snf4 compatible: Adds restriction enzyme site HindIII on C-terminus and Snf4 tail on N-terminus. <br>
+
-
                                  Expected fragment length: 769 bp.<br>
+
-
                                  5. Amplification of cfp-snf1 compatible: Adds restriction enzyme site SpeI on N-terminus and Snf1 tail on C-terminus.<br>
+
-
                                  Expected fragment length: 757 bp.<br>
+
-
                                  K2. Control K2.<br>
+
-
                                  <br>
+
-
                                  Julia, Katarina and Malin<br>
+
-
                                  <strong>Verified PCR 1-5:</strong> measured concentration by nanodrop  and checked if the fragments were the correct length by running on a 2 %  agarose gel (8 µl sample+2 µl loading buffer, Fermentas FastRuler Low  Range).<br>
+
-
                                  PCR product 1, 2, 3 and 5 worked and gave expected fragment lengths.  Number 4 however did not work.  Only a fragment around 80 bp was  visible, probably a primer dimer.<br>
+
-
                                  <br>
+
-
                                  <strong>Concentrations:</strong><br>
+
-
                                  1. 443.9 ng/µl<br>
+
-
                                  2. 371.0 ng/µl<br>
+
-
                                  3. 361.7 ng/µl<br>
+
-
                                  4. --<br>
+
-
                                  5. 377.6 ng/µl<br>
+
-
                                  A. 451.1 ng/µl (snf1)<br>
+
-
                                  B. 438.7 ng/µl (snf4)<br>
+
-
                                  C. 408.9 ng/µl (SAMS) <br>
+
-
                                  1-5 and A-C were diluted to 1 ng/µl.<br>
+
-
                                  <br>
+
-
                                  <strong>Ran PCR of I, II, III, IV and 4.</strong><br>
+
-
                                  4.2.  Amplification of cfp-Snf4 compatible: Adds restriction enzyme site  HindIII on C-terminus and Snf4 tail on N-terminus. Expected fragment  length: 769 bp. (second try)<br>
+
-
                                  I. PCR fusion of cfp and SAMS: Adds BamHI on N-terminus of cfp and gives a SAMS C-terminus. Expected fragment length: 768 bp.<br>
+
-
                                  II. PCR fusion of yfp and Snf4 1: Adds BamHI on N-terminus of yfp and removes stop codon. Expected fragment length: 1694 bp.<br>
+
-
                                  III. PCR fusion of yfp and Snf4 2: Adds BamHI on N-terminus of yfp and  adds HindIII on C-terminus of Snf4 plus adds stop codon. Expected  fragment length: 1708 bp. Final Product!<br>
+
-
                                  IV. PCR fusion of cfp and Snf1: Adds SpeI on N-terminus of cfp and PacI  on C-terminus of Snf1 and keeps the stop codon. Expected fragment  length: 2643 bp.<br>
+
-
                                  K3. Control<br>
+
-
                                  <br>
+
-
                                  Karl and Malin<br>
+
-
                                  Transferred diploid cells of snf1Δ mutants to two sporulation plates and incubated in 30° C. (For 2-7 days)<br>
+
-
                                </p></li>
+
-
                             
+
-
                                <li><p><strong>2010-07-13</strong><br>
+
-
                                  Julia and  Lokesh<br>
+
-
                                  Ran the gel  with products of our first three PCRs along with the  templates we made for the  two fluorescent proteins. We also prepared  the sample solutions of DNA  templates with appropriate concentration  (1ng/µl) for the next PCR reactions.<br>
+
-
                                  <strong>Verified</strong> the PCR products and the digested miniprepped    EYFP and ECFP by running an agarose gel which showed good results.  Expected  fragment lengths were received. The SAMS-peptide however was  too short to be  visible on the gel (67 bp).<br>
+
-
                                  <strong>Prepared for PCR 1-5. </strong>The miniprepped ECFP and EYFP  were  diluted to a stock concentration of 1 ng/ ml to use for the PCR.   The concentrations of the first PCR product  were measured using  nanodrop and then diluted to 1 ng/ml.<br>
+
-
                                  snf1: 20.5 ng/ml<br>
+
-
                                  snf4: 122.6 ng/ml<br>
+
-
                                  SAMS: 114.9 ng/ml<br>
+
-
                                </p>
+
-
                                <p> Karl and  Adnan<br>
+
-
                                  We made  duplicates of the haploid Snf4∆ and wildtype strains on new  plates in case the  original plate would be contaminated. Prepared a  sporulation medium that was  autoclaved and poured into empty growth  plates to stay in room temperature over  night.</p>
+
-
                                  <br></li>
+
-
        <li>  <p> <strong>2010-07-12</strong><br>
+
-
                                  Katarina  and Julia<br>
+
-
                                    <strong>Amplification of genomic SNF1 DNA by PCR. </strong>All  primers were resuspended in  mq-water according to the volume on the  lable, the final concentration 100 pg/ml. The primers were then diluted  10x  and the SAMS-peptide templates were diluted 1000x. PCR reactions  were run  amplifying subunits Snf1 and Snf4 and also the SAMS-peptide.  YPD medium and  agar plates were prepared (yeast).<br>
+
-
                                  </p>
+
-
                                  <p> Adnan and  Kemal<br>
+
-
                                <strong>Miniprep purification </strong>of EYFP and ECFP plasmides from  overnight culture. The concentrations were measured using Nanodrop:<br>
+
-
                                  ECFP1: 99 ng/ml<br>
+
-
                                  ECFP2: 89.9<br>
+
-
                                  EYFP1: 104, 7 <br>
+
-
                                  EYFP2: 172,5<br>
+
-
                                  The  concentrations were verified with  electrophoresis. The plasmids were cut with  restriction enzymes EcoRI  and Pst1.<br>
+
-
                                  ECFP, plasmid pSB1AK3, fragment sizes 750 bp and 3200 bp.<br>
+
-
                                  EYFP, plasmid pSB1A2, fragment sizes 750 bp and 2050 bp.</p></li>
+
-
                                <br>
+
-
                                <li><p><strong>2010-07-11</strong><br>
+
-
                                    Katarina<br>
+
-
                                    Made  overnight cultures with colony 3  and 4 plasmid (not single cultures) and twp  each of EYFP and ECFP.
+
-
                                    </p>
+
-
                                </li>
+
-
                                <br>
+
-
                                <li><p><strong>2010-07-10</strong><br>
+
-
                                    Julia<br>
+
-
                                    Moved  plated bacteria from incubator to refrigerator. <br>
+
-
                                  </p>
+
-
                                </li>
+
-
                                <li>
+
-
                                    <p><strong>2010-07-09</strong><br>
+
-
                                      Malin and  Julia<br>
+
-
                                      The SOC  medium was autoclaved.  Competent E.coli cells were transformed with our FPs  through heat shock  and plated on petri dishes. 20 new LB-amp plates were made.<br>
+
-
                                     
+
-
                                      Adnan and  Katarina<br>
+
-
                                      Picked up  primers at Chalmers and  checked if they were ok. The concentration of the  genomic DNA (SNF1  gene) were measured. Prepared a master mix so that the  PCR-reactions  could be started Monday morning.</p>
+
-
                                  <br>
+
-
                                </li>
+
-
                                <li> 
+
-
                                  <p><strong>2010-07-08</strong><br>
+
-
                                    Malin,  Julia and Peidi<br>
+
-
                                    The two  BioBricks EYFP and ECFP were  extracted from the kit plates and stored in the  freezer. A SOC-medium  was prepared.<br>
+
-
                                  </p>
+
-
                                </li>
+
-
                                <li>
+
-
                                  <p><strong>2010-07-07 </strong><br>
+
-
                                    Adnan,  Katarina and Julia<br>
+
-
                                    As the gels  from Tuesday showed  ambigous results we decided to redo the miniprep of the  plamids. We  used the overnight preparations of the two unused colonies (3 &amp;  4).  The miniprep went smoothly except for major difficulties regarding    untrustworthy pipettes. The pipettes marked with green tape are the ones  to use  from now on together with the tips with the same markings! A  gel was molded and  this time we decided to use a full gel to get a  better resolution.  The results on the gel were very good.  The plasmids  had clearly been cut in one or  two places and the fragment sizes seem  to correspond to the expected sizes.<br>
+
-
                                  </p>
+
-
                                </li>
+
-
                                <li> 
+
-
                                  <p><strong>2010-07-06</strong><br>
+
-
                                    Malin, Adnan and Peidi<br>
+
-
                                    Colony 1 and 2 were picked to do a  miniprep.  The plasmids were extracted and a plasmid integrity check was  performed. By  cutting with BamHI, one cut were made to get the size of  the plasmid. Then we  cut with ClaI which is a two-cutter that gives a  fragment that is 600 bp and  one that is 6800. The gel which should  confirm this was not satisfactory since  the 600 band was missing.<br>
+
-
                                  </p>
+
-
                                </li>
+
-
                                <li>
+
-
                                  <p><strong>2010-07-05</strong><br>
+
-
                                    Malin and Katarina<br>
+
-
                                    An overnight culture were made from 4  different  colonies of E.coli containing the plasmid pSB-GM1.</p>
+
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Latest revision as of 21:02, 20 September 2010

Chalmers University of Technology

Team
FUSS

We are a team of 8 students from Chalmers University of Technology who will represent Gothenburg, SWEDEN in this year’s iGEM competition. We have started with a promising idea that combines the cutting edge technologies available in the field of Synthetic Biology. Our research basically aims to constructing a reporter mechanism for cellular stress in yeast by tagging protein and peptides affiliated with the stress activated SNF1 complex with fluorescent markers.

Chalmers University of Technology

Chalmers in central Göteborg on the Swedish West Coast

Chalmers is a university of technology in which research and teaching are conducted on a broad front within technology, natural science and architecture. Our inspiration lies in the joy of discovery and the desire to learn. Underlying everything we do is a wish to contribute to sustainable development both in Sweden and world-wide.

Our research work deals ultimately with improving people’s conditions, and we often cross traditional boundaries in order to solve the problems of the future. Chalmers has become strong within several areas of science, and some of our research leads its field internationally. We wish in particular to develop and strengthen our research in the fields of bioscience, information technology, environmental science and nanotechnology.

Chalmers is a pioneer in Europe in the field of start-up companies, and has built up an efficient system of innovation in order to find applications in research into the formation of companies.

Chalmers has great aspirations to develop continually by opening doors to the outside world and by thinking innovatively. We have a close and productive collaboration with business and society at large. In this way we reinforce our own development, at the same time as we contribute to the growth of society.

Chalmers was founded in 1829 as a result of a donation from the director of the Swedish East India Company, William Chalmers. His motto, and ours, is:

Avancez!

Supervisors
Goutham Vemuri
supervisor

 

 

 

Per Sunnerhagen
supervisor

 

 

 

Marcus Wilhelmsson
supervisor

 

 

   
Students
Malin Linden
student
 
Age: 24
Nationality: Swedish
Academic background: B sc. Biotecnology
Current studies: Exchange year at University of Minnesota, MS Biotecnology
   

 

Katarina Risö
student
 
Age: 24
Nationality: Swedish
Academic background: B sc. Chemical Engineering
Current studies: MS Chemistry and Biosciences
   

 

Katarina Risö
student
 
Age: 24
Nationality: Swedish
Academic background: B sc. Chemical Engineering
Current studies: MS Chemistry and Biosciences
   

 

Adnan Kadic
student
 
Age: 24
Nationality: Bosnian
Academic background: B sc. Biotecnology
Current studies: MS Chemistry and Biosciences
   

 

Lokeshwaran Manoharan
student
 
Age: 23
Nationality: Indian
Academic background: B.Tech Ind Biotechnology
Current studies: MS Bioinformatics
   

 

Kemal Sanli
student
 
Age: 24
Nationality: Turkish
Academic background: B sc. Molecular Biology and Genetics
Current studies: MS Bioinformatics
   

 

Karl Almén Burman
student
 
Age: 23
Nationality: Swedish
Academic background: B sc. Biotechnology
Current studies: MS Biotechnology
   

 

Julia Fransson
student
 
Age: 22
Nationality: Swedish
Academic background: B sc. Biotechnology
Current studies: Exchange year through Unitech International
   

 

   

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Chalmers University of Technology, Gothenburg, SWEDEN