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| - | __NOTOC__
| + | <html> |
| - | ==Calendar== | + | <style type="text/css"> |
| - | {|{| class="toccolours" style="float:{{{float|left}}}; width:{{{width|200px}}}; margin-left:1em; text-align:center;" | + | <!-- |
| | + | h2 { |
| | + | font-weight:bold; |
| | + | } |
| | + | |
| | + | #contentSub { |
| | + | display:none; |
| | + | } |
| | + | |
| | + | #siteSub { |
| | + | display:none; |
| | + | } |
| | + | |
| | + | #search-controls { |
| | + | display:none; |
| | + | } |
| | + | |
| | + | .firstHeading { |
| | + | display:none; |
| | + | } |
| | + | |
| | + | #search-controls { |
| | + | display:none; |
| | + | } |
| | + | |
| | + | #footer-box { |
| | + | |
| | + | } |
| | + | |
| | + | #top-section { |
| | + | height:25px; |
| | + | width:1000px; |
| | + | color: blue; |
| | + | background-color: transparent; |
| | + | background-image: url(https://static.igem.org/mediawiki/2010/f/f1/Osaka_top_section.jpg) |
| | + | } |
| | + | |
| | + | #p-logo { |
| | + | display:none; |
| | + | } |
| | + | |
| | + | #menubar { |
| | + | top:0px; |
| | + | } |
| | + | |
| | + | #globalWrapper { |
| | + | background-color: transparent; |
| | + | background-image: url(https://static.igem.org/mediawiki/2010/a/af/Osaka_sand2.jpg); |
| | + | padding-bottom: 20px; |
| | + | padding-left:0px; |
| | + | padding-right:0px; |
| | + | } |
| | + | } |
| | + | |
| | + | #content { |
| | + | position: relative; |
| | + | width:1000px; |
| | + | padding:0px; |
| | + | } |
| | + | .wrap{ |
| | + | |
| | + | } |
| | + | .top{ |
| | + | width:1000px; |
| | + | background-color:black; |
| | + | } |
| | + | .logo{ width:160px; |
| | + | float:left; |
| | + | padding:0; |
| | + | margin:0; |
| | + | background-color:#0342ff; |
| | + | |
| | + | } |
| | + | .banner{ |
| | + | width:840px; |
| | + | float:left; |
| | + | padding:0; |
| | + | margin:0; |
| | + | background-color:black; |
| | + | |
| | + | } |
| | + | |
| | + | .bottom{ |
| | + | width:1000px;} |
| | + | .right{ |
| | + | width:840px; |
| | + | float:right; |
| | + | } |
| | + | .contents2{ |
| | + | width:820px; |
| | + | float:right; |
| | + | padding:0 10px; |
| | + | margin:0; |
| | + | background-color:white; |
| | + | } |
| | + | .twitter{ |
| | + | width:250px; |
| | + | float:right; |
| | + | padding:0; |
| | + | margin:0; |
| | + | background-color:white; |
| | + | } |
| | + | .contents{ |
| | + | width:750px; |
| | + | float:left; |
| | + | padding:0 10px; |
| | + | margin:0; |
| | + | background-color:white; |
| | + | } |
| | + | |
| | + | .menu{ |
| | + | width:160px; |
| | + | float:left; |
| | + | background-color:white; |
| | + | padding:0; |
| | + | } |
| | + | .clear{ |
| | + | clear:both;} |
| | + | .clear hr{ |
| | + | display:none;} |
| | + | .bold { |
| | + | font-weight:bold; |
| | + | } |
| | + | --> |
| | + | </style> |
| | + | </head> |
| | + | |
| | + | |
| | + | <body> |
| | + | |
| | + | <div class="wrap"> |
| | + | <div class="top"> |
| | + | <div class="banner"> |
| | + | <img src="https://static.igem.org/mediawiki/2010/d/d6/Osaka_banner3.jpg" width="1000" height="250" alt="iGEM Osaka banner" usemap="#banner_map"> |
| | + | |
| | + | <map name="banner_map"> |
| | + | <area shape="rect" coords="540,180,750,220" href="https://2010.igem.org" alt="iGEM 2010 home"/> |
| | + | <area shape="rect" coords="430,230,750,270" href="http://www.osaka-u.ac.jp/en" alt="Osaka University"/> |
| | + | <area shape="circle" coords="100,200,85" href="https://2010.igem.org/Team:Osaka" alt="iGEM Osaka home"/> |
| | + | </map> |
| | + | |
| | + | </div><!-- banner --> |
| | + | <div class="clear"><hr></div> |
| | + | </div><!-- top --> |
| | + | |
| | + | <div class="bottom"> |
| | + | <div class="menu"> |
| | + | </html> |
| | + | {{Osaka_menu}} |
| | + | <html> |
| | + | </div> |
| | + | |
| | + | |
| | + | |
| | + | |
| | + | <html> |
| | + | <div class="right"> |
| | + | <div class="contents"> |
| | + | <div style="margin:40px 0"> |
| | + | </html> |
| | + | ==Notebook== |
| | + | {| class="toccolours" style="float:{{{float|left}}}; width:{{{width|240px}}}; text-align:center;" |
| | |- style="background:{{{color1|#ccccff}}};" | | |- style="background:{{{color1|#ccccff}}};" |
| - | |colspan="8"|{{{header|July}}} | + | |colspan="8"|{{{header|July / August}}} |
| | |- | | |- |
| | |- style="background:{{{color2|#eeeeff}}};" | | |- style="background:{{{color2|#eeeeff}}};" |
| - | |wodth=100px|Week | + | |width=100px|Week |
| | |width="14%"|<span style="color:red;">S</span> | | |width="14%"|<span style="color:red;">S</span> |
| | |width="14%"|M | | |width="14%"|M |
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| | |width="14%"|F | | |width="14%"|F |
| | |width="14%"|<span style="color:deepskyblue;">S</span> | | |width="14%"|<span style="color:deepskyblue;">S</span> |
| - | |-
| |
| - | |colspan ="5"|
| |
| - | |<span style={{{s1|"color:{{{c1|inherit}}};"}}}>1</span>
| |
| - | |<span style={{{s2|"color:{{{c2|inherit}}};"}}}>2</span>
| |
| - | |<span style={{{s3|"color:{{{c3|deepskyblue}}};"}}}>3</span>
| |
| - | |-
| |
| - | |<span style></span>
| |
| - | |<span style={{{s4|"color:{{{c4|red}}};"}}}>4</span>
| |
| - | |<span style={{{s5|"color:{{{c5|inherit}}};"}}}>5</span>
| |
| - | |<span style={{{s6|"color:{{{c6|inherit}}};"}}}>6</span>
| |
| - | |<span style={{{s7|"color:{{{c7|inherit}}};"}}}>7</span>
| |
| - | |<span style={{{s8|"color:{{{c8|inherit}}};"}}}>8</span>
| |
| - | |<span style={{{s9|"color:{{{c9|inherit}}};"}}}>9</span>
| |
| - | |<span style={{{s10|"color:{{{c10|deepskyblue}}};"}}}>10</span>
| |
| - | |-
| |
| - | |<span style></span>
| |
| - | |<span style={{{s11|"color:{{{c11|red}}};"}}}>11</span>
| |
| - | |<span style={{{s12|"color:{{{c12|inherit}}};"}}}>12</span>
| |
| - | |<span style={{{s13|"color:{{{c13|inherit}}};"}}}>13</span>
| |
| - | |<span style={{{s14|"color:{{{c14|inherit}}};"}}}>14</span>
| |
| - | |<span style={{{s15|"color:{{{c15|inherit}}};"}}}>15</span>
| |
| - | |<span style={{{s16|"color:{{{c16|inherit}}};"}}}>16</span>
| |
| - | |<span style={{{s17|"color:{{{c17|deepskyblue}}};"}}}>17</span>
| |
| - | |-
| |
| - | |<span style></span>
| |
| - | |<span style={{{s18|"color:{{{c18|red}}};"}}}>18</span>
| |
| - | |<span style={{{s19|"color:{{{c19|inherit}}};"}}}>19</span>
| |
| - | |<span style={{{s20|"color:{{{c20|inherit}}};"}}}>20</span>
| |
| - | |<span style={{{s21|"color:{{{c21|inherit}}};"}}}>21</span>
| |
| - | |<span style={{{s22|"color:{{{c22|inherit}}};"}}}>22</span>
| |
| - | |<span style={{{s23|"color:{{{c23|inherit}}};"}}}>23</span>
| |
| - | |<span style={{{s24|"color:{{{c24|deepskyblue}}};"}}}>24</span>
| |
| | |- | | |- |
| | |<span style>[[Team:Osaka/week1|week1]]</span> | | |<span style>[[Team:Osaka/week1|week1]]</span> |
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| | |<span style={{{s30|"color:{{{c30|inherit}}};"}}}>30</span> | | |<span style={{{s30|"color:{{{c30|inherit}}};"}}}>30</span> |
| | |[[Team:Osaka/week1#July 31 (Sat)|31]]<span style={{{s31|"color:{{{c31|deepskyblue}}};"}}}></span> | | |[[Team:Osaka/week1#July 31 (Sat)|31]]<span style={{{s31|"color:{{{c31|deepskyblue}}};"}}}></span> |
| - | |-
| |
| - | |- style="background:{{{color3|#eeeeff}}};"
| |
| - | |colspan="7"|{{{footer|}}}
| |
| - | |}<noinclude>
| |
| - | {| class="toccolours" style="float:{{{float|left}}}; width:{{{width|200px}}}; margin-left:1em; text-align:center;"
| |
| - | |- style="background:{{{color1|#ccccff}}};"
| |
| - | |colspan="8"|{{{header|August}}}
| |
| - | |-
| |
| - | |- style="background:{{{color2|#eeeeff}}};"
| |
| - | |width=100px|Week
| |
| - | |width="14%"|<span style="color:red;">S</span>
| |
| - | |width="14%"|M
| |
| - | |width="14%"|T
| |
| - | |width="14%"|W
| |
| - | |width="14%"|T
| |
| - | |width="14%"|F
| |
| - | |width="14%"|<span style="color:deepskyblue;">S</span>
| |
| | |- | | |- |
| | |<span style>[[Team:Osaka/week2|week2]]</span> | | |<span style>[[Team:Osaka/week2|week2]]</span> |
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| | |colspan="7"|{{{footer|}}} | | |colspan="7"|{{{footer|}}} |
| | |}<noinclude> | | |}<noinclude> |
| - | {| class="toccolours" style="float:{{{float|left}}}; width:{{{width|200px}}}; margin-left:1em; text-align:center;" | + | {| class="toccolours" style="float:{{{float|left}}}; width:{{{width|240px}}}; text-align:center;" |
| | |- style="background:{{{color1|#ccccff}}};" | | |- style="background:{{{color1|#ccccff}}};" |
| | |colspan="8"|{{{header|September}}} | | |colspan="8"|{{{header|September}}} |
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| | |<span style>[[Team:Osaka/week7|week7]]</span> | | |<span style>[[Team:Osaka/week7|week7]]</span> |
| | |[[Team:Osaka/week7#September 5 (Sun)|5]]<span style={{{s5|"color:{{{c5|red}}};"}}}></span> | | |[[Team:Osaka/week7#September 5 (Sun)|5]]<span style={{{s5|"color:{{{c5|red}}};"}}}></span> |
| - | |[[Team:Osaka/week7#September 6 (Sun)|5]]<span style={{{s6|"color:{{{c6|inherit}}};"}}}></span> | + | |[[Team:Osaka/week7#September 6 (Mon)|6]]<span style={{{s6|"color:{{{c6|inherit}}};"}}}></span> |
| - | |[[Team:Osaka/week7#September 7 (Sun)|5]]<span style={{{s7|"color:{{{c7|inherit}}};"}}}></span> | + | |[[Team:Osaka/week7#September 7 (Tue)|7]]<span style={{{s7|"color:{{{c7|inherit}}};"}}}></span> |
| - | |[[Team:Osaka/week7#September 8 (Sun)|5]]<span style={{{s8|"color:{{{c8|inherit}}};"}}}></span> | + | |[[Team:Osaka/week7#September 8 (Wed)|8]]<span style={{{s8|"color:{{{c8|inherit}}};"}}}></span> |
| - | |[[Team:Osaka/week7#September 9 (Sun)|5]]<span style={{{s9|"color:{{{c9|inherit}}};"}}}></span> | + | |[[Team:Osaka/week7#September 9 (Thu)|9]]<span style={{{s9|"color:{{{c9|inherit}}};"}}}></span> |
| - | |[[Team:Osaka/week7#September 10 (Sun)|5]]<span style={{{s10|"color:{{{c10|inherit}}};"}}}></span> | + | |[[Team:Osaka/week7#September 10 (Fri)|10]]<span style={{{s10|"color:{{{c10|inherit}}};"}}}></span> |
| - | |[[Team:Osaka/week7#September 11 (Sun)|5]]<span style={{{s11|"color:{{{c11|deepskyblue}}};"}}}></span> | + | |[[Team:Osaka/week7#September 11 (Sat)|11]]<span style={{{s11|"color:{{{c11|deepskyblue}}};"}}}></span> |
| | |- | | |- |
| | |<span style>[[Team:Osaka/week8|week8]]</span> | | |<span style>[[Team:Osaka/week8|week8]]</span> |
| - | |<span style={{{s12|"color:{{{c12|red}}};"}}}>12</span> | + | |[[Team:Osaka/week8#September 12 (Sun)|12]]<span style={{{s12|"color:{{{c12|red}}};"}}}></span> |
| - | |<span style={{{s13|"color:{{{c13|inherit}}};"}}}>13</span> | + | |[[Team:Osaka/week8#September 13 (Mon)|13]]<span style={{{s13|"color:{{{c13|inherit}}};"}}}></span> |
| - | |<span style={{{s14|"color:{{{c14|inherit}}};"}}}>14</span> | + | |[[Team:Osaka/week8#September 14 (Tue)|14]]<span style={{{s14|"color:{{{c14|inherit}}};"}}}></span> |
| - | |<span style={{{s15|"color:{{{c15|inherit}}};"}}}>15</span> | + | |[[Team:Osaka/week8#September 15 (Wed)|15]]<span style={{{s15|"color:{{{c15|inherit}}};"}}}></span> |
| - | |<span style={{{s16|"color:{{{c16|inherit}}};"}}}>16</span> | + | |[[Team:Osaka/week8#September 16 (Thu)|16]]<span style={{{s16|"color:{{{c16|inherit}}};"}}}></span> |
| - | |<span style={{{s17|"color:{{{c17|inherit}}};"}}}>17</span> | + | |[[Team:Osaka/week8#September 17 (Fri)|17]]<span style={{{s17|"color:{{{c17|inherit}}};"}}}></span> |
| - | |<span style={{{s18|"color:{{{c18|deepskyblue}}};"}}}>18</span> | + | |[[Team:Osaka/week8#September 18 (Sat)|18]]<span style={{{s18|"color:{{{c18|deepskyblue}}};"}}}></span> |
| | |- | | |- |
| | |<span style>[[Team:Osaka/week9|week9]]</span> | | |<span style>[[Team:Osaka/week9|week9]]</span> |
| - | |<span style={{{s19|"color:{{{c19|red}}};"}}}>19</span> | + | |[[Team:Osaka/week9#September 19 (Sun)|19]]<span style={{{s19|"color:{{{c19|red}}};"}}}></span> |
| - | |<span style={{{s20|"color:{{{c20|inherit}}};"}}}>20</span> | + | |[[Team:Osaka/week9#September 20 (Mon)|20]]<span style={{{s20|"color:{{{c20|inherit}}};"}}}></span> |
| - | |<span style={{{s21|"color:{{{c21|inherit}}};"}}}>21</span> | + | |[[Team:Osaka/week9#September 21 (Tue)|21]]<span style={{{s21|"color:{{{c21|inherit}}};"}}}></span> |
| - | |<span style={{{s22|"color:{{{c22|inherit}}};"}}}>22</span> | + | |[[Team:Osaka/week9#September 22 (Wed)|22]]<span style={{{s22|"color:{{{c22|inherit}}};"}}}></span> |
| - | |<span style={{{s23|"color:{{{c23|inherit}}};"}}}>23</span> | + | |[[Team:Osaka/week9#September 23 (Thu)|23]]<span style={{{s23|"color:{{{c23|inherit}}};"}}}></span> |
| - | |<span style={{{s24|"color:{{{c24|inherit}}};"}}}>24</span> | + | |[[Team:Osaka/week9#September 24 (Fri)|24]]<span style={{{s24|"color:{{{c24|inherit}}};"}}}></span> |
| - | |<span style={{{s25|"color:{{{c25|deepskyblue}}};"}}}>25</span> | + | |[[Team:Osaka/week9#September 25 (Sat)|25]]<span style={{{s25|"color:{{{c25|deepskyblue}}};"}}}></span> |
| | |- | | |- |
| | |<span style>[[Team:Osaka/week10|week10]]</span> | | |<span style>[[Team:Osaka/week10|week10]]</span> |
| - | |<span style={{{s26|"color:{{{c26|red}}};"}}}>26</span> | + | |[[Team:Osaka/week10#September 26 (Sun)|26]]<span style={{{s26|"color:{{{c26|red}}};"}}}></span> |
| - | |<span style={{{s27|"color:{{{c27|inherit}}};"}}}>27</span> | + | |[[Team:Osaka/week10#September 27 (Mon)|27]]<span style={{{s27|"color:{{{c27|inherit}}};"}}}></span> |
| - | |<span style={{{s28|"color:{{{c28|inherit}}};"}}}>28</span> | + | |[[Team:Osaka/week10#September 28 (Tue)|28]]<span style={{{s28|"color:{{{c28|inherit}}};"}}}></span> |
| - | |<span style={{{s29|"color:{{{c29|inherit}}};"}}}>29</span> | + | |[[Team:Osaka/week10#September 29 (Wed)|29]]<span style={{{s29|"color:{{{c29|inherit}}};"}}}></span> |
| - | |<span style={{{s30|"color:{{{c30|inherit}}};"}}}>30</span> | + | |[[Team:Osaka/week10#September 30 (Thu)|30]]<span style={{{s30|"color:{{{c30|inherit}}};"}}}></span> |
| | |colspan="2"| | | |colspan="2"| |
| | |- style="background:{{{color3|#eeeeff}}};" | | |- style="background:{{{color3|#eeeeff}}};" |
| | |colspan="7"|{{{footer|}}} | | |colspan="7"|{{{footer|}}} |
| | |}<noinclude> | | |}<noinclude> |
| - | {| class="toccolours" style="float:{{{float|left}}}; width:{{{width|150px}}}; margin-left:1em; text-align:center;" | + | {| class="toccolours" style="float:{{{float|left}}}; width:{{{width|240px}}}; text-align:center;" |
| | |- style="background:{{{color1|#ccccff}}};" | | |- style="background:{{{color1|#ccccff}}};" |
| - | |colspan="7"|{{{header|October}}} | + | |colspan="8"|{{{header|October}}} |
| | |- | | |- |
| | |- style="background:{{{color2|#eeeeff}}};" | | |- style="background:{{{color2|#eeeeff}}};" |
| | + | |width=100px|Week |
| | |width="14%"|<span style="color:red;">S</span> | | |width="14%"|<span style="color:red;">S</span> |
| | |width="14%"|M | | |width="14%"|M |
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| | |width="14%"|<span style="color:deepskyblue;">S</span> | | |width="14%"|<span style="color:deepskyblue;">S</span> |
| | |- | | |- |
| - | |colspan ="5"| | + | |<span style>[[Team:Osaka/week10|week10]]</span> |
| - | |<span style={{{s1|"color:{{{c1|inherit}}};"}}}>1</span> | + | |<span style></span> |
| - | |<span style={{{s2|"color:{{{c2|deepskyblue}}};"}}}>2</span> | + | |<span style></span> |
| | + | |<span style></span> |
| | + | |<span style></span> |
| | + | |<span style></span> |
| | + | |[[Team:Osaka/week10#October 1 (Fri)|1]]<span style={{{s1|"color:{{{c1|inherit}}};"}}}></span> |
| | + | |[[Team:Osaka/week10#October 2 (Sat)|2]]<span style={{{s2|"color:{{{c2|deepskyblue}}};"}}}></span> |
| | |- | | |- |
| - | |<span style={{{s3|"color:{{{c3|red}}};"}}}>3</span> | + | |<span style>[[Team:Osaka/week11|week11]]</span> |
| - | |<span style={{{s4|"color:{{{c4|inherit}}};"}}}>4</span> | + | |[[Team:Osaka/week11#October 3 (Sun)|3]]<span style={{{s3|"color:{{{c3|red}}};"}}}></span> |
| - | |<span style={{{s5|"color:{{{c5|inherit}}};"}}}>5</span> | + | |[[Team:Osaka/week11#October 4 (Mon)|4]]<span style={{{s4|"color:{{{c4|inherit}}};"}}}></span> |
| - | |<span style={{{s6|"color:{{{c6|inherit}}};"}}}>6</span> | + | |[[Team:Osaka/week11#October 5 (Tue)|5]]<span style={{{s5|"color:{{{c5|inherit}}};"}}}></span> |
| - | |<span style={{{s7|"color:{{{c7|inherit}}};"}}}>7</span> | + | |[[Team:Osaka/week11#October 6 (Wed)|6]]<span style={{{s6|"color:{{{c6|inherit}}};"}}}></span> |
| - | |<span style={{{s8|"color:{{{c8|inherit}}};"}}}>8</span> | + | |[[Team:Osaka/week11#October 7 (Thu)|7]]<span style={{{s7|"color:{{{c7|inherit}}};"}}}></span> |
| - | |<span style={{{s9|"color:{{{c9|deepskyblue}}};"}}}>9</span> | + | |[[Team:Osaka/week11#October 8 (Fri)|8]]<span style={{{s8|"color:{{{c8|inherit}}};"}}}></span> |
| | + | |[[Team:Osaka/week11#October 9 (Sat)|9]]<span style={{{s9|"color:{{{c9|deepskyblue}}};"}}}></span> |
| | |- | | |- |
| - | |<span style={{{s10|"color:{{{c10|red}}};"}}}>10</span> | + | |<span style>[[Team:Osaka/week12|week12]]</span> |
| - | |<span style={{{s11|"color:{{{c11|inherit}}};"}}}>11</span> | + | |[[Team:Osaka/week12#October 10 (Sun)|10]]<span style={{{s10|"color:{{{c10|red}}};"}}}></span> |
| - | |<span style={{{s12|"color:{{{c12|inherit}}};"}}}>12</span> | + | |[[Team:Osaka/week12#October 11 (Mon)|11]]<span style={{{s11|"color:{{{c11|inherit}}};"}}}></span> |
| - | |<span style={{{s13|"color:{{{c13|inherit}}};"}}}>13</span> | + | |[[Team:Osaka/week12#October 12 (Tue)|12]]<span style={{{s12|"color:{{{c12|inherit}}};"}}}></span> |
| - | |<span style={{{s14|"color:{{{c14|inherit}}};"}}}>14</span> | + | |[[Team:Osaka/week12#October 13 (Wed)|13]]<span style={{{s13|"color:{{{c13|inherit}}};"}}}></span> |
| - | |<span style={{{s15|"color:{{{c15|inherit}}};"}}}>15</span> | + | |[[Team:Osaka/week12#October 14 (Thu)|14]]<span style={{{s14|"color:{{{c14|inherit}}};"}}}></span> |
| - | |<span style={{{s16|"color:{{{c16|deepskyblue}}};"}}}>16</span> | + | |[[Team:Osaka/week12#October 15 (Fri)|15]]<span style={{{s15|"color:{{{c15|inherit}}};"}}}></span> |
| | + | |[[Team:Osaka/week12#October 16 (Sat)|16]]<span style={{{s16|"color:{{{c16|deepskyblue}}};"}}}></span> |
| | |- | | |- |
| - | |<span style={{{s17|"color:{{{c17|red}}};"}}}>17</span> | + | |<span style>[[Team:Osaka/week13|week13]]</span> |
| - | |<span style={{{s18|"color:{{{c18|inherit}}};"}}}>18</span> | + | |[[Team:Osaka/week13#October 17 (Sun)|17]]<span style={{{s17|"color:{{{c17|red}}};"}}}></span> |
| - | |<span style={{{s19|"color:{{{c19|inherit}}};"}}}>19</span> | + | |[[Team:Osaka/week13#October 18 (Mon)|18]]<span style={{{s18|"color:{{{c18|inherit}}};"}}}></span> |
| - | |<span style={{{s20|"color:{{{c20|inherit}}};"}}}>20</span> | + | |[[Team:Osaka/week13#October 19 (Tue)|19]]<span style={{{s19|"color:{{{c19|inherit}}};"}}}></span> |
| - | |<span style={{{s21|"color:{{{c21|inherit}}};"}}}>21</span> | + | |[[Team:Osaka/week13#October 20 (WEd)|20]]<span style={{{s20|"color:{{{c20|inherit}}};"}}}></span> |
| - | |<span style={{{s22|"color:{{{c22|inherit}}};"}}}>22</span> | + | |[[Team:Osaka/week13#October 21 (Thu)|21]]<span style={{{s21|"color:{{{c21|inherit}}};"}}}></span> |
| - | |<span style={{{s23|"color:{{{c23|deepskyblue}}};"}}}>23</span> | + | |[[Team:Osaka/week13#October 22 (Fri)|22]]<span style={{{s22|"color:{{{c22|inherit}}};"}}}></span> |
| | + | |[[Team:Osaka/week13#October 23 (Sat)|23]]<span style={{{s23|"color:{{{c23|deepskyblue}}};"}}}></span> |
| | |- | | |- |
| - | |<span style={{{s24|"color:{{{c24|red}}};"}}}>24</span> | + | |<span style>[[Team:Osaka/week14|week14]]</span> |
| - | |<span style={{{s25|"color:{{{c25|inherit}}};"}}}>25</span> | + | |[[Team:Osaka/week14#October 24 (Sun)|24]]<span style={{{s24|"color:{{{c24|red}}};"}}}></span> |
| - | |<span style={{{s26|"color:{{{c26|inherit}}};"}}}>26</span> | + | |[[Team:Osaka/week14#October 25 (Mon)|25]]<span style={{{s25|"color:{{{c25|inherit}}};"}}}></span> |
| - | |<span style={{{s27|"color:{{{c27|inherit}}};"}}}>27</span> | + | |[[Team:Osaka/week14#October 26 (Tue)|26]]<span style={{{s26|"color:{{{c26|inherit}}};"}}}></span> |
| | + | |[[Team:Osaka/week14#October 27 (Wed)|27]]<span style={{{s27|"color:{{{c27|inherit}}};"}}}></span> |
| | |<span style={{{s28|"color:{{{c28|inherit}}};"}}}>28</span> | | |<span style={{{s28|"color:{{{c28|inherit}}};"}}}>28</span> |
| | |<span style={{{s29|"color:{{{c29|inherit}}};"}}}>29</span> | | |<span style={{{s29|"color:{{{c29|inherit}}};"}}}>29</span> |
| | |<span style={{{s30|"color:{{{c30|deepskyblue}}};"}}}>30</span> | | |<span style={{{s30|"color:{{{c30|deepskyblue}}};"}}}>30</span> |
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| | + | |<span style>[[Team:Osaka/week15|week15]]</span> |
| | |[[{{{prefix|}}}10.31{{{suffix|}}}|<span style={{{s31|"color:{{{c31|red}}};"}}}>31</span>]] | | |[[{{{prefix|}}}10.31{{{suffix|}}}|<span style={{{s31|"color:{{{c31|red}}};"}}}>31</span>]] |
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| - | ==Notebook==
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| - |
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| - | =
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| - | =
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| - | ===September 12 (Sun)===
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| - | # Miniprep of Fcex, 006, 007, 010, 011
| |
| - | # Restriction digests of 006, 007, 010, 011 with EcoRI, SpeI; Fcex with XbaI, PstI
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| - | # Gel electrophoresis of digests
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| - | #* (RESULTS)
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| - | # Ligation for 3A assembly
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| - | #* 014: 011 as upstream, 1-13D as downstream, 1-3A as vector
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| - | # Transformation of 014 using 2μl ligation product with 50μl competent cells
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| - | # Moved yesterday's transformation plates (012, 013) to 4°C refrigerator ''no RFP expression from vector plasmids at all?''
| |
| - | # Transfer of A01, A02, A03 (previously transformed) to solution culture
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| - |
| |
| - | ===September 13 (Mon)===
| |
| - | # Miniprep of A01, A03; restriction digest with EcoRI followed by electrophoresis
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| - | #* (RESULTS?)
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| - | # PCR test
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| - | #* re-cloning of beta-glucosidase from F1 (received from Edinburgh team) into Silver standard-compatible format
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| - | #* cloning of pgsC from A01
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| - | #* ''note: primers were misdesigned but PCR performed anyway to confirm whether sequences were complementary, and to identify PCR parameters''
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| - | # Gel electrophoresis of PCR products
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| - | #* (RESULTS?)
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| - | # Gel purification of PCR product from F1
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| - | # Restriction digest of PCR product (F1) and 1-5A, both with EcoRI, SpeI
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| - | # Gel electrophoresis of PCR product from A01
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| - | #* band visible around 400~500bp, which was correct length -> PCR conditions identified!
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| - | # Transformation of A01, A02, A03
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| - | # Transfer of 012, 013, 014 to solution culture
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| - | #* ''RFP expressed from vector plasmids on 012, 013 plates during refrigeration period, so selection of colonies with inserts now possible''
| |
| - |
| |
| - | ===September 14 (Tue)===
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| - | # Miniprep of A02, 012, 013, 014
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| - | # Restriction digests
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| - | #* 012, 013, 014 with EcoRI, PstI
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| - | #* A02 with EcoRI
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| - | # Gel electrophoresis of digests
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| - | #* A02 was discarded (bad size?)
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| - | #* 012~014: digestions repeated with 1μl each of EcoRI, PstI (previous digestions were 0.5μl each)
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| - | #* gel electrophoresis of repeat digestions again showed bad sizes for all parts
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| - | #** 1-13D terminator part is bad? -> cut check of 1-13D
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| - | #** failure to inactivate restriction enzymes before ligations? -> re-ligation
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| - | # 1-13D cut check with XbaI, PstI
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| - | # Re-ligation (3A assembly)
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| - | #* previously digested 009, 008, 011 inactivated at 80°C for 20min before ligation
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| - | #* 012: 009 as upstream, 1-13D as downstream, 1-3A as vector (same as on 9/11)
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| - | #* 013: 008 as upstream, 1-13D as downstream, 1-3A as vector (same as on 9/11)
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| - | #* 014: 011 as upstream, 1-13D as downstream, 1-3A as vector (same as on 9/12)
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| - | # Transformation of ligated products (012~014)
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| - | # Gel electrophoresis of yesterday's restriction digests of 1-5A and PCR product from F1 followed by their ligation (1-5A as vector, PCR product as insert)
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| - | # Transfer of A01, A02, A03 to LB liquid culture medium supplanted with 25μl of 2M glucose to prevent leaky expression from Lac promoter (may be affecting growth)
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| - | # Received additional cellulase parts from Karita-sensei: Cel5, Xyn10, Cel44, Man26, Cel8
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| - | #* plasmid DNA resuspended in 10μl MiliQ water each
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| - | #* transformation with 2μl DNA solution
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| - |
| |
| - | ===September 15 (Wed)===
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| - | # Miniprep A02, A03 (''E. coli failed to grow in A01'')
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| - | # Cut check of A02, A03 with EcoRI
| |
| - | #* A02 is ok
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| - | # Transformation of 3-22G
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| - | #* (INFO?)
| |
| - | # PCR cloning of pgsC from A02
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| - | # Colony check & transfer of cellulase parts from Karita-sensei to solution culture
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| - | # Preparation of lysis buffer for yeast genome DNA extraction according to [insert reference]
| |
| - | {|
| |
| - | !Component!!Volume Added!!Final Concentration
| |
| - | |-
| |
| - | |Triton X-100||100μl||2%
| |
| - | |-
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| - | |10%SDS||500μl||1%
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| - | |-
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| - | |5M NaCl||100μl||100mM
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| - | |-
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| - | |20mM Tris-HCl(ph8.0)||2.5ml||10mM
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| - | |-
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| - | |0.5M EDTA(ph8.0)||10μl||1mM
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| - | |-
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| - | |dH2O||1790μl
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| - | |-
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| - | |'''TOTAL'''||5ml
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| - | |}
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| - | # PCR to clone pgsA, pgsB by 'Megaprimer' method using NEB Phusion polymerase
| |
| - |
| |
| - | ===September 16 (Thu)===
| |
| - | # Gel electrophoresis to verify yesterday's PCR results
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| - | #* pgsA megaprimer: 750bp -> OK
| |
| - | #* pgsB megaprimer: 170bp -> OK
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| - | #* pgsC: 450bp -> very faint band?
| |
| - | # Gel extraction of pgsA, pgsB megaprimers
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| - | # 2nd step of megaprimer PCR for pgsA, pgsB
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| - | #* failure; possible causes:
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| - | #** short annealing step?
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| - | #** low denaturation temperature?
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| - | #** mistakes in procedure?
| |
| - | # PCR
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| - | #* pgsC using correct (redesigned) primers
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| - | #* pgsA, B megaprimer 2nd step repeat using reaction mix composition modified from OpenWetWare
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| - | #* gel purification of PCR products
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| - | # Miniprep of BglX (temporary designation for part cloned from F1 with faulty primer), Cel5, Cel8, Cel44, Man26, Xyn10
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| - | # Cut check of miniprepped parts with XbaI, PstI
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| - | # Yeast genome DNA extraction (detailed protocol will be provided elsewhere) according to [reference article]
| |
| - | # Gel electrophoresis
| |
| - | #* yeast genome DNA
| |
| - | #* PCR products
| |
| - | #** pgsC -> OK
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| - | # Restriction digest
| |
| - | #* pgsC (PCR product) with EcoRI, PstI
| |
| - | #* 1-1C, 1-3A, 1-5A (vectors) with EcoRI, SpeI
| |
| - |
| |
| - | ===September 17 (Fri)===
| |
| - | # PCR cloning of ADH1 terminator from yeast genome DNA
| |
| - | #* 4 simultaneous attempts with varying template concentration, thermocycle settings & timing of polymerase addition (before or after initial denaturation)
| |
| - | #* gel electrophoresis showed no PCR product obtained for any of the reactions -> ''annealing temperatures were too stringent?''
| |
| - | # PCR of ADH1 terminator (repeat)
| |
| - | #* annealing temperature was lowered from 70°C to 60°C
| |
| - | #* PCR failed again -> ''possible RNA contamination?''
| |
| - | # PCR of ADH1 terminator (2nd repeat)
| |
| - | #* genomic DNA treated with RNase (0.1μl added to 1μl genome DNA; incubation at 37°C for 15min) before using as template
| |
| - | # Gel electrophoresis of yesterday's digests: pgsC, 1-1C, 1-3A, 1-5A
| |
| - | # Miniprep of 013 and 3-22G
| |
| - | #* 012, 014 culture solutions have turned red -> picked-up colonies had RFP-carrying vectors, not ligated plasmids; discarded!
| |
| - | #Cut check of 3-22G with XbaI, PstI; 013 with EcoRI, PstI
| |
| - | #* (RESULTS?)
| |
| - | # Ligations
| |
| - | #* 018: 3A assembly with 004 as upstream, Fcex as downstream, 1-1C as vector
| |
| - | #* 019: pgsC as insert, 1-1C as vector (''cut at?'')
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| - | # PCR cloning of XynA CBM, pgsA
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| - | # Colony pick-up & transfer to solution culture: 006, 007 (9/10 ligation/transformation) ''more plasmids needed?''
| |
| - | # Transformation of 004, 005, 008, 009, 018, 019
| |
| - |
| |
| - | ===September 18 (Sat)===
| |
| - | # PCR cloning of pgsA by overlap extension ''megaprimer method doesn't seem to work so well''
| |
| - | # Gel electrophoresis of PCR product (after overlap step) of pgsA
| |
| - | #* correct band seems to be obtained
| |
| - | # Gel extraction followed by restriction digest of PCR product with EcoRI, PstI
| |
| - | # Miniprep of 006, 007
| |
| - | # Restriction digest with EcoRI, PstI followed by gel electrophoresis
| |
| - | # PCR of pgsB (1st fragment for overlap extension)
| |
| - | #* ''tried 2 times but couldn't get amplification product!''
| |
| - | # Transfer to solution culture: 004, 005, 008, 009, 018, 019
| |
| - |
| |
| - | ===September 19 (Sun)===
| |
| - | # PCR of pgsB (repeat)
| |
| - | #* again, no product :(
| |
| - | # PCR of pgsB (4th attempt, including yesterday's)
| |
| - | #* no product
| |
| - | # Miniprep of 004, 005, 008, 009, 018, 019
| |
| - | # Restriction digest of 018 with EcoRI, SpeI; 019 with XbaI, PstI
| |
| - | # Gel electrophoresis of pgsA, 004, 005, 008, 009, 018, 019
| |
| - | #* apart from 018 & 019, all are parts digested before; used here to compare with new parts 018, 019
| |
| - | #* (RESULTS?)
| |
| - | #* problem with 019?
| |
| - | # PCR of pgsB 1st fragment (5th attempt)
| |
| - | #* (RESULTS?)
| |
| - | # PCR of pgsB - generation of 2nd fragment (170bp) for overlap extension
| |
| - | # PCR of pgsB - generation of 3rd fragment (1000bp) for overlap extension
| |
| - | # PCR: Phusion activity check using BglX as template & the primers that generated it
| |
| - | # PCR: pgsB template check using the outermost primers
| |
| - |
| |
| - | ===September 20 (Mon)===
| |
| - | # Transfer to solution culture: 004, 005, 006, 007, 008 ,009 ,019
| |
| - | # PCR: Phusion polymerase & template checks (repeat of 9/19?)
| |
| - | #* positive control (BglX) was amplified -> Phusion polymerase seems to be working ok
| |
| - | #* problem with template? primer? thermocycle settings?
| |
| - | # PCR: primer check
| |
| - | #* pair of primers for each overlap segment were tested
| |
| - | #* (RESULTS?)
| |
| - | # Digestion of pgsA (PCR product) and 1-1C with EcoRI, PstI
| |
| - | # Ligation to make new part 020: pgsA as insert, 1-1C as vector
| |
| - | # Transformation of ligation product
| |
| - | # PCR of pgsB 1st fragment (''n-th repeat??'')
| |
| - | * '''Note: MANY MANY rounds of PCR carried out today; due to time constraints they are not described here in detail'''
| |
| - |
| |
| - | ===September 21 (Tue)===
| |
| - | # Miniprep of yesterday's cultures: 004, 005, 006, 007, 008 ,009; 019 was discarded (turned red)
| |
| - | # Restriction digest of miniprepped parts with EcoRI, SpeI
| |
| - | # Gel electrophoresis of digested parts 004, 005, 006, 007, 008, 009, 019 (?)
| |
| - | #* 005 - OK
| |
| - | #* 014 - no band visible
| |
| - | #* 019 - bad length - ''repeat ligation''
| |
| - | #* 006, 007 - bad lengths; ''repeat colony pick-up & culture?''
| |
| - | # PCR to synthesize C-terminal half of pgsB (overlap extension method continued)
| |
| - | #* band of correct size obtained!
| |
| - | # PCR cloning of Man26B, CelB
| |
| - | #* gel run failed to turn up bands; repeat with lower annealing temp
| |
| - | #* repeat run succeeded!
| |
| - | # Inoculated YPD liquid culture medium with yeast
| |
| - | # New part 020 (contains pgsA) transferred to solution culture
| |
| - | # PCR of pgsB (final step - extension of overlapping fragments)
| |
| - | # Gel electrophoresis to extract PCR products (Man26, CelB) as well as 1-5A for plasmid backbone
| |
| - |
| |
| - | ===September 22 (Wed)===
| |
| - | # Gel electrophoresis of 1-5A, 1-3A, 1-1C, PCR product (pgsB) followed by extraction/purification
| |
| - | # Restriction digest of extracted parts with EcoRI, SpeI
| |
| - | # Miniprep of 020
| |
| - | # Restriction digest of 020 with XbaI, PstI (for gel run to check & further assembly)
| |
| - | # Restriction digest of pgsC, pgsA with EcoRI, SpeI
| |
| - | # Gel electrophoresis of all digested parts above
| |
| - | #* ''020 was bad; repeat ligation?''
| |
| - | # Transfer of 006, 007 to solution culture (pick up from new colonies?)
| |
| - | # Ligations
| |
| - | #* 019: pgsC (PCR product) into 1-1C vector
| |
| - | #* 020: pgsA (PCR product) into 1-1C vector
| |
| - | #* 021: pgsB (PCR product) into 1-1C vector
| |
| - | #* all using PCR products purified today
| |
| - | # Transformation of above ligation products
| |
| - | # Extraction of genome DNA from yeast cultured yesterday
| |
| - | # PCR cloning of yeast parts from genomic DNA
| |
| - | #* ADH1 terminator
| |
| - | #* ADH2 promoter
| |
| - | #* CYC1 terminator
| |
| - | #* ENO2 promoter
| |
| - | #* SUC2 leader sequence
| |
| - |
| |
| - | ===September 23 (Thu)===
| |
| - | # Gel electrophoresis of yesterday's PCR products followed by extraction
| |
| - | # Restriction digest of PCR products with EcoRI, SpeI
| |
| - | #* ENO2, ADH2 incorrect length -> repeat
| |
| - | # PCR cloning of ENO2 promoter, ADH2 promoter, glr (glutamate racemase)
| |
| - | # Gel electrophoresis of crude PCR product, extraction & purification from gel
| |
| - | #* ENO2 promoter, ADH2 promoter, glr obtained!
| |
| - | # PCR cloning of CelB, Man26B, Cel44A (Cel44A: internal mutations needed; 1st step of overlap extension fragment generation)
| |
| - | # Gel electrophoresis of CelB, Man26B, Cel44A PCR products
| |
| - | #* (RESULTS?)
| |
| - | # Miniprep of 006, 007 followed by restriction digest with EcoRI, SpeI
| |
| - | # Gel electrophoresis of 006, 007
| |
| - | #* (RESULTS?)
| |
| - | # Transfer to solution culture: 019, 020
| |
| - | #* no white/non-RFP colonies on 021 (pgsB) plate
| |
| - | #** insert (PCR product) was not digested properly?
| |
| - | #** problem with gel purification?
| |
| - |
| |
| - | ===September 24 (Fri)===
| |
| - | # Inoculation of Karita-sensei's cellulase parts into fresh LB (need more plasmid); also, 024 (beta-glucosidase)
| |
| - | # Extraction from iGEM distribution plates:
| |
| - | {|
| |
| - | !ID!!Part Name!!Resistance!!Description
| |
| - | |-
| |
| - | |1-6N||<bbpart>BBa_</bbpart>||A,K||T7 promoter
| |
| - | |-
| |
| - | |2-2F||<bbpart>BBa_</bbpart>||A||T7 polymerase
| |
| - | |-
| |
| - | |1-6I||<bbpart>BBa_</bbpart>||A||tetracycline-repressible promoter
| |
| - | |}
| |
| - | # PCR purification of yesterday's Cel44A
| |
| - | # Miniprep of 019, 020
| |
| - | # Restriction digest of 019, 020 with XbaI, PstI
| |
| - | #* inserts of correct lengths obtained!
| |
| - | # Ligations for 3A assembly
| |
| - | #* 004: 001 as upstream, 1-2J as downstream, 1-3A as vector
| |
| - | #* 005: 001 as upstream, 2-22P as downstream, 1-3A as vector
| |
| - | #* pgsB 10xHC 1-3A ''???''
| |
| - | # Transformation of ligation products
| |
| - | # PCR cloning (repeat) of Man26, CelB
| |
| - | # Gel electrophoresis
| |
| - | #* (RESULTS?)
| |
| - |
| |
| - | ===September 25 (Sat)===
| |
| - | # Miniprep of yesterday's solution cultures (cellulase parts-containing plasmids from Karita-sensei)
| |
| - | # Restriction digest of miniprepped plasmid DNA with XbaI, PstI (''??? these are not biobrick plasmids!'') & gel electrophoresis
| |
| - | # Transformation of miniprepped parts
| |
| - | # Restriction digests
| |
| - | #* 001 with EcoRI, PstI
| |
| - | #* K1 (''??'') with EcoRI, SpeI
| |
| - | # Ligations to transfer 001, K1 into 1-1C (Amp-resistance) vector -> designated as 001-2
| |
| - | #* 025: xylanase (K1) in 1-1C vector
| |
| - | # PCR cloning of CelB
| |
| - | # Transformation of 001-2, 025
| |
| - |
| |
| - |
| |
| - | ===September 26 (Sun)===
| |
| - | # Transfer to culture solution (yesterday's transformations)
| |
| - | # Miniprep of 1-6N, 2-2F, 1-6I, 021 ''faint hint of red detected''
| |
| - | # Restriction digest
| |
| - | #* 1-6N, 1-6I with EcoRI, SpeI
| |
| - | #* 2-2F, 021 with XbaI, PstI
| |
| - | # Gel electrophoresis
| |
| - | #* (RESULTS?)
| |
| - | # Miniprep of parts moved to solution culture this morning
| |
| - | # Cut check with XbaI, PstI <- ''??these are not biobrick plasmids!''
| |
| - | #* Cel8 - ok
| |
| - | #* Cel44 - ??
| |
| - | #* Man26 - ok
| |
| - | #* Xyn10 - ??
| |
| - | # Ligations of 9/23 restriction digests (ADH1, ADH2, CYC1, ENO2, SUC2, glr) to 1-1C plasmid backbone digested at E, S sites
| |
| - | #* 026 - ADH1 terminator
| |
| - | #* 027 - ADH2 promoter
| |
| - | #* 028 - CYC1 terminator
| |
| - | #* 029 - ENO2 promoter
| |
| - | #* 030 - SUC2 leader sequence
| |
| - | #* 031 - glr (glutamate racemase)
| |
| - | # Transformation of ligation products
| |
| - | # Transfer of yesterday's transformations to solution culture: 001-2, 025
| |
| - |
| |
| - | ===September 27 (Mon)===
| |
| - | # PCR of Man26, CelB
| |
| - | # PCR of Man48 (??), CBM from XynAcc
| |
| - | # Restriction digests
| |
| - | #* 1-2M with EcoRI, SpeI for assembly later
| |
| - | #* Cel8, Cel44, Cel5 with EcoRI; Xyn10 with PstI (''for checking?'')
| |
| - | #* 001-2 with EcoRI, SpeI
| |
| - | #* 025 with XbaI, PstI
| |
| - | # Gel electrophoresis
| |
| - | # Ligations
| |
| - | #* 032: 1-2M as upstream, 019 as downstream, 1-3A as vector
| |
| - | #* 033: 1-2M as upstream, 020 as downstream, 1-3A as vector
| |
| - | #* 004: 001-2 as upstream, 1-2J as downstream, 1-3A as vector (''remake using 001-2'')
| |
| - | #* 005: 001-2 as upstream, 2-22P as downstream, 1-3A as vector (<nowiki>" "</nowiki>)
| |
| - | # Transformation of ligation products
| |
| - | # Transfer to solution culture: 026, 027, 028, 029, 030, 031
| |
| - |
| |
| - | ===September 28 (Tue)===
| |
| - | # Miniprep of 026, 027, 028, 030, 031
| |
| - | # Restriction digest
| |
| - | #* 026, 028, 031 with XbaI, PstI
| |
| - | #* 027, 030 with EcoRI, SpeI
| |
| - | # Gel electrophoresis
| |
| - | #* (RESULTS?)
| |
| - | # Ligation: repeat assemblies of 004, 005, 032, 033 yesterday
| |
| - | # Transformation of ligation products as well as 3-2P
| |
| - | {|
| |
| - | !ID!!Part Name!!Resistance!!Description
| |
| - | |-
| |
| - | |3-2P||<bbpart>BBa_</bbpart>||A||??
| |
| - | |}
| |
| - |
| |
| - | ===September 29 (Wed)===
| |
| - | # Transfer to solution culture 025, 026, 027, 028, 030, 031
| |
| - | # Ligations (repeat of 9/26 with different dilution of 1-1C)
| |
| - | #* 026 - ADH1 terminator
| |
| - | #* 027 - ADH2 promoter
| |
| - | #* 028 - CYC1 terminator
| |
| - | #* 029 - ENO2 promoter
| |
| - | #* 030 - SUC2 leader sequence
| |
| - | #* 031 - glr (glutamate racemase)
| |
| - | # Transformation of 025, 026, 027, 028, 030, 031 (''repeat just in case; previous plates almost all red colonies; cannot pick up colony with correct insert?'')
| |
| - |
| |
| - | # PCR
| |
| - | #* pgsB - cloning from plasmid
| |
| - | #* pgsB - amplification from 021
| |
| - | #* Man28 - amplification from previous product
| |
| - | # PCR purification of products
| |
| - |
| |
| - | # PCR of CelB, XynA-CBM
| |
| - |
| |
| - | # Miniprep of cultures inoculated this morning (total culture time: 12hr)
| |
| - | #* 025 -> ok (colorless)
| |
| - | #* 027 -> turned red; discarded
| |
| - | #* others: ok?
| |
| - | # Restriction digest of 026, 028, 031 with XbaI, PstI
| |
| - | # Transfer to solution culture (9/28 transformations of 004, 005, 032, 033, 3-2P)
| |
| - | # Gel electrophoresis of PCR products
| |
| - |
| |
| - | # Restriction digests:
| |
| - | #* pgsB, Man (??) with EcoRI, SpeI
| |
| - | #* today's miniprepped plasmids with EcoRI
| |
| - | #* 1-1C with EcoRI, SpeI
| |
| - |
| |
| - | ===September 30 (Thu)===
| |
| - | # PCR cloning
| |
| - | #* CM10
| |
| - | #* CelB (repeat)
| |
| - | #* ADH1 terminator, CYC1 terminator, SUC2 leader sequence from yeast genome (repeat)
| |
| - | #* glr (repeat)
| |
| - | # PCR purification: CM10, XynA-CBM
| |
| - | # Restriction digest of all above PCR products with EcoRI, SpeI
| |
| - | # Gel electrophoresis
| |
| - |
| |
| - | # Ligation of PCR products to 1-1C backbone
| |
| - | #* 021: pgsB
| |
| - | #* 025: XynA-CBM
| |
| - | #* 026: ADH1 terminator
| |
| - | #* 028: CYC1 terminator
| |
| - | #* 031: glr
| |
| - | #* 034: Man
| |
| - | #* 035: CM10
| |
| - | # Transformation of ligation products
| |
| - |
| |
| - | # PCR cloning of CelB, SUC2, ECO2, ADH2 from genomic DNA
| |
| - | # Gel electrophoresis
| |
| - | #* (RESULTS?)
| |
| - |
| |
| - | # Miniprep of 004, 005, 032, 033, 3-2P (9/29 solution culture)
| |
| - | # Restriction digest of miniprepped parts
| |
| - | #* 004, 005, 032 with EcoRI, SpeI (same as earlier today)
| |
| - | #* 033, 3-2P (same as on 9/29)
| |
| - | # Gel electrophoresis
| |
| - | #* (RESULTS?)
| |
| - |
| |
| - | ===TO CHECK/CONFIRM===
| |
| - | * 'K1' (mentioned on 9/250 -> where did it come from?
| |
| - | * 9/25 - miniprepped parts (inoculated 9/24) were plasmids from karita sensei? biobrick parts pcr-cloned on 9/23? why where these parts transformed immediately after miniprep?
| |
| - | * 'CelB, Cel44A' etc vs 'Cel5, Cel8, Cel44' which nomenclature refers to what (received plasmids, pcr product?)
| |
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