Team:Osaka/Notebook

From 2010.igem.org

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==Notebook==
+
<html>
 +
<style type="text/css">
 +
<!--
 +
h2 {
 +
font-weight:bold;
 +
}
-
===July 29 (Thu)===
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#contentSub {
-
Attendance: Torigata, Takino, Teoh, Yasumoto, Kakuda, Saka, Tamura
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display:none;
-
# Safety lecture for junior members.
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}
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# Preparation of LB agar plates (26 Amp, 25 Kan, 25 Cam).
+
-
===July 31 (Sat)===
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#siteSub {
-
Attendance: Miyatake, Hirayama, Torigata, Teoh, Tadashi, Yasumoto, Kakuda, Saka
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display:none;
-
# Meeting
+
}
-
#* Summer project schedule
+
-
#* List of genes to clone
+
-
===August 2 (Mon)===
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#search-controls {
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Attendance: Torigata, Takino, Teoh, Tadashi, Yasumoto, Saka
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display:none;
-
# Culture medium preparation
+
}
-
#* LB agar plates (49 antibiotic-less plates)
+
-
#* LB liquid medium (500 ml)
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-
# Competent cells preparation - Nojima Method
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-
#* SOB medium (MgCl2 not yet added) -> stored at 4˚C
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-
#* TB buffer -> stored at 4˚C
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-
#* Single-colony streaking of E. coli (strain DH5α) on 2 LB agar plates -> 37˚C incubation o/n
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-
Tomorrow we shall prepare the remainder of the reagents and inoculate SOB with o/n-incubated E. coli.
+
-
===August 3 (Tue)===
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.firstHeading {
-
Attendance: Nakamura, Kakuda, Saka, Yasumoto, Teoh
+
display:none;
-
# Competent cells preparation (continued)
+
}
-
#* Preparation of glucose solution for making SOC medium.
+
-
#* Inoculation of pre-culture SOB from o/n-incubated agar plate colonies.
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-
#* (Night) Transfer from pre-culture to growth culture.
+
-
===August 4 (Wed)===
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#search-controls {
-
Attendance: Nakamura, Saka, Kakuda, Yasumoto, Torigata, Teoh
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display:none;
-
# OD measurements throughout the day till required OD (0.3~0.7) was obtained.
+
}
-
# Completion of competent cells according to protocol.
+
-
===August 5 (Thu)===
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#footer-box {
-
Attendance: Nakamura, Yasumoto, Saka, Kakuda, Takino
+
-
# Transformation of Registry parts:
+
-
{|
+
-
!ID!!Part Name!!Resistance!!Description
+
-
|-
+
-
|2-20J||<bbpart>BBa_K118023</bbpart>||A||''C. fermi'' endocellulase Cen A coding
+
-
|-
+
-
|2-20H||<bbpart>BBa_K118022</bbpart>||A||''C. fermi'' exocellulase Cex coding
+
-
|-
+
-
|1-2M||<bbpart>BBa_B0034</bbpart>||A||RBS
+
-
|-
+
-
|1-13D||<bbpart>BBa_B0010</bbpart>||A||terminator
+
-
|-
+
-
|1-1D||<bbpart>BBa_R0010</bbpart>||A||promoter
+
-
|-
+
-
|1-18F||<bbpart>BBa_E1010</bbpart>||K||RFP coding
+
-
|}
+
-
Note: 'ID' is an internal identifier used by wet work members to simplify labeling etc. In the case of iGEM distribution parts it usually refers to the plate location.
+
-
===August 6 (Fri)===
+
}
-
Attendance: Nakamura, Saka, Yasumoto, Takino, Teoh
+
-
# Colony check
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-
#* All transformed cells produced colonies!
+
-
#* Non-transformed cells (negative controls) did not grow on Amp, Kan or Cam plates -> confirmed lack of natural antibiotic resistance
+
-
# Colonies transferred to 3ml LB growth medium & incubated o/n at 37˚C
+
-
===August 7 (Sat)===
+
#top-section {
-
Attendance: Nakamura, Saka, Yasumoto, Takino
+
height:25px;
-
# Miniprep of o/n-incubated colonies (2-20J, 2-20H, 1-2M, 1-13D, 1-1D, 1-18F) using Sigma-Aldrich Plasmid Miniprep Kit
+
width:1000px;
-
# Transformation of construction plasmids
+
color: blue;
-
{|
+
background-color: transparent;
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!ID!!Part Name!!Resistance!!Description
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background-image: url(https://static.igem.org/mediawiki/2010/f/f1/Osaka_top_section.jpg)
-
|-
+
}
-
|1-1C||<bbpart>pSB1A3</bbpart>||A||construction plasmid containing mRFP coding device (<bbpart>BBa_J04450</bbpart>)
+
-
|-
+
-
|1-3A||<bbpart>pSB1C3</bbpart>||C||<nowiki>(" ")</nowiki>
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-
|-
+
-
|1-5A||<bbpart>pSB1K3</bbpart>||K||<nowiki>(" ")</nowiki>
+
-
|}
+
-
# Meeting
+
-
===August 8 (Sun)===
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#p-logo {
-
Attendance: Nakamura, Yasumoto
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display:none;
-
# Colony check
+
}
-
#*All parts successfully transformed
+
-
# Transfer to LB culture medium
+
-
===August 9 (Mon)===
+
#menubar {
-
# Miniprep of 1-1C
+
top:0px;
-
# Restriction digests of 2-20H, 2-20J, 1-1D, 1-18F, 1-1C, 1-13D, 1-2M
+
}
-
#* (WHICH ENZYMES?)
+
-
# Gel electrophoresis of digests ("cut check")
+
-
#* 2-20H, 2-20J, 1-1C -> OK
+
-
#* 1-1D, 1-18F, 1-13D, 1-2M -> not cut (single band, MW approx. equal to vector + insert)
+
-
# Transfer of 1-3A, 1-5A colonies to solution culture (repeat)
+
-
#*''Yesterday's inoculated culture mediums contained the wrong antibiotics!''
+
-
# Transformation of <bbpart>BBa_I13521</bbpart>, <bbpart>BBa_I13522</bbpart>, <bbpart>BBa_I13600</bbpart>, <bbpart>BBa_K204031</bbpart>, <bbpart>BBa_K204051</bbpart>, <bbpart>BBa_K204032</bbpart> from last year's stock
+
-
===August 10 (Tue)===
+
#globalWrapper {
-
# Miniprep of 1-3A, 1-5A
+
background-color: transparent;
-
# Restriction digests of 1-3A, 1-5A
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background-image: url(https://static.igem.org/mediawiki/2010/a/af/Osaka_sand2.jpg);
-
# Gel electrophoresis
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padding-bottom: 20px;
-
## 1st run: 1-3A, 1-5A (newly miniprepped), 1-1D, 1-18F, 1-13D, 1-2M (repeat)
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padding-left:0px;
-
##* 1-3A, 1-5A -> OK; others -> not cut (''again'')
+
padding-right:0px;
-
## 2nd run: 1-1D, 1-18F, 1-13D, 1-2M (2nd repeat)
+
}
-
##* all parts not cut
+
}
-
===August 11 (Wed)===
+
#content {
-
# Miniprep of last year's parts transformed on Monday
+
position: relative;
-
# Gel electrophoresis of 1-1D, 1-18F, 1-13D, 1-2M (3rd repeat!!!)
+
width:1000px;
-
#* all 4 not cut... AGAIN
+
padding:0px;
-
#* so far all restriction digests involving XbaI seem to have failed; problem with enzyme stock?
+
}
-
#* will try with different set of restriction enzymes next week
+
.wrap{
-
# Sent miniprepped last year's parts to ECUST team in Shanghai, China
+
 +
}
 +
.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;
 +
 +
}
-
===August 16 (Mon)===
+
.bottom{
-
# Restriction digests
+
width:1000px;}
-
#* 1-13D,　1-2M,　1-1D,　1-18F with EcoRI, PstI to check for point mutation(s) affecting XbaI site
+
.right{
-
#* 2-20J with XbaI, PstI to check/confirm XbaI activity
+
width:840px;
-
# Gel electrophoresis of digests
+
float:right;
-
#* (RESULTS?)
+
}
-
# Colony pick-up & transfer to solution culture (repeat; 2 each): 1-13D,　1-2M,　1-1D,　1-18F
+
.contents2{
-
#* 3ml LB liquid medium; 3μl Amp or 0.6μl Kan solution added
+
        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;
 +
}
-
===August 17 (Tue)===
+
.menu{
-
# Mini-prep of 1-13D,　1-2M,　1-1D,　1-18F parts inoculated yesterday (2 of each)
+
width:160px;
-
# 'Cut check' (restriction digest + gel electrophoresis to confirm insert) of the miniprepped parts:
+
float:left;
-
#* 1-1D, 1-18F with EcoRI, SpeI
+
background-color:white;
-
#* 1-13D, 1-2M with XbaI, PstI
+
        padding:0;
-
#* (RESULTS?)
+
}
-
# Transformation of secretion tag parts using 25μl of competent cells each
+
.clear{
-
{|
+
clear:both;}
-
!ID!!Part Name!!Resistance!!Description
+
.clear hr{
-
|-
+
display:none;}
-
|2-22P||<bbpart>BBa_K103006</bbpart>||A||OmpA outer membrane protein + linker
+
.bold {
-
|-
+
      font-weight:bold;
-
|1-2J||<bbpart>BBa_J32015</bbpart>||A,K||PelB leader sequence
+
}
-
|}
+
-->
 +
</style>
 +
</head>
-
===August 18 (Wed)===
 
-
iGEM Japan Meet-Up in Kyoto
 
-
Attendance: Nakamura, Yasumoto, Saka, Kakuda
 
-
===August 19 (Thu)===
+
<body>
-
# Transfer of 2-22P, 1-2J to solution culture
+
-
# Gel electrophoresis of digests from 'cut check' products from Tuesday
+
-
#* repeat run, but each digest together with undigested plasmid DNA)
+
-
#* 2% agarose gel instead of the usual 1%
+
-
#* (RESULTS?)
+
-
# Gel electrophoresis of 1-1D digest only
+
-
#* (RESULT?)
+
-
# Multiple restriction digests of 1-1D to check for problems at restriction sites
+
-
#* tried the following: EcoRI only; SpeI only; EcoRI + SpeI
+
-
# Night: miniprep of 2-22P, 1-2J inoculated in the morning
+
-
===August 20 (Fri)===
+
<div class="wrap">
-
# Gel electrophoresis of 1-1D and its digests
+
<div class="top">
-
#* (RESULTS?)
+
    <div class="banner">
-
# 'Cut check' of parts miniprepped the night before
+
  <img src="https://static.igem.org/mediawiki/2010/d/d6/Osaka_banner3.jpg" width="1000" height="250" alt="iGEM Osaka banner" usemap="#banner_map">
-
#* both 2-22P & 1-2J cut with XbaI, PstI
+
-
#* enzyme inactivation at 80℃, 20min
+
-
#* (RESULTS?)
+
-
# Restriction digest of 2-20J (WHICH ENZYMES?)
+
-
# Ligation according to 3A assembly method: 2-20J or 2-20H + 2-22P or 1-2J; 1-3A as vector
+
-
#* reaction mixture: 2μl of each plasmid, 2μl ligase buffer, 1μl T4 DNA ligase, water to make 20μl total
+
-
#* reaction at room temperature for 10min; ligase inactivation at 80℃ for 20min
+
-
# Transformation of ligated parts using 50μl of competent cells each; 2μl ligation product; 150μl Chloramphenicol spread on agar plates before inoculation with pre-incubation mix
+
-
===August 21 (Sat)===
+
<map name="banner_map">
-
# Transfer of white (non-RFP) colonies from yesterday's transformation to 3ml LB liquid medium added with 1μl Cam solution
+
  <area shape="rect" coords="540,180,750,220" href="https://2010.igem.org" alt="iGEM 2010 home"/>
-
#* ''we later realized that the upstream and downstream parts had been mixed up in these ligations, so the ligation products (and these corresponding solution cultures) were discarded''
+
  <area shape="rect" coords="430,230,750,270" href="http://www.osaka-u.ac.jp/en" alt="Osaka University"/>
-
# 3A assembly ligation: 1-1D as upstream, 1-2M as downstream, 1-3A as vector
+
  <area shape="circle" coords="100,200,85" href="https://2010.igem.org/Team:Osaka" alt="iGEM Osaka home"/>
-
#* ligation product designated as <1>
+
</map>
-
#* same ligation mix composition as yesterday's
+
 
-
# Transformation of <1> with pre-incubation for 1.5hr instead of 1hr, transformation mix inoculated onto Chloramphenicol plate
+
  </div><!-- banner -->
 +
  <div class="clear"><hr></div>
 +
</div><!-- top -->
 +
 
 +
<div class="bottom">
 +
  <div class="menu">
 +
</html>
 +
{{Osaka_menu}}
 +
<html>
 +
  </div>
-
===August 22 (Sun)===
+
 
-
# Transfer of <1> to culture solution; incubation at 30℃ (''why??'')
+
 
-
#* Transformation of the following parts:
+
 
-
{|
+
<html>
-
!ID!!Part Name!!Resistance!!Description
+
<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}}};"
 +
|colspan="8"|{{{header|July / August}}}
|-
|-
-
|2-4A||<bbpart>BBa_J63005</bbpart>||A||yeast ADH1 promoter
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|- 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>
|-
|-
-
|3-11J||<bbpart>BBa_K098987</bbpart>||K||temperature sensitive cI inducible system with GFP reporter and low promoter
+
|<span style>[[Team:Osaka/week1|week1]]</span>
 +
|<span style={{{s25|"color:{{{c25|red}}};"}}}>25</span>
 +
|<span style={{{s26|"color:{{{c26|inherit}}};"}}}>26</span>
 +
|<span style={{{s27|"color:{{{c27|inherit}}};"}}}>27</span>
 +
|<span style={{{s28|"color:{{{c28|inherit}}};"}}}>28</span>
 +
|[[Team:Osaka/week1#July 29 (Thu)|29]]<span style={{{s29|"color:{{{c29|inherit}}};"}}}></span>
 +
|<span style={{{s30|"color:{{{c30|inherit}}};"}}}>30</span>
 +
|[[Team:Osaka/week1#July 31 (Sat)|31]]<span style={{{s31|"color:{{{c31|deepskyblue}}};"}}}></span>
|-
|-
-
|2-20A||<bbpart>BBa_I729006</bbpart>||A||AHL reporter and aiia device
+
|<span style>[[Team:Osaka/week2|week2]]</span>
 +
|<span style={{{s1|"color:{{{c1|red}}};"}}}>1</span>
 +
|[[Team:Osaka/week2#August 2 (Mon)|2]]<span style={{{s2|"color:{{{c2|inherit}}};"}}}></span>
 +
|[[Team:Osaka/week2#August 3 (Tue)|3]]<span style={{{s3|"color:{{{c3|inherit}}};"}}}></span>
 +
|[[Team:Osaka/week2#August 4 (Wed)|4]]<span style={{{s4|"color:{{{c4|inherit}}};"}}}></span>
 +
|[[Team:Osaka/week2#August 5 (Thu)|5]]<span style={{{s5|"color:{{{c5|inherit}}};"}}}></span>
 +
|[[Team:Osaka/week2#August 6 (Fri)|6]]<span style={{{s6|"color:{{{c6|inherit}}};"}}}></span>
 +
|[[Team:Osaka/week2#August 7 (Sat)|7]]<span style={{{s7|"color:{{{c7|deepskyblue}}};"}}}></span>
|-
|-
-
|F1||N/A||A||beta-galactosidase from Edinburgh team
+
|<span style>[[Team:Osaka/week3|week3]]</span>
 +
|[[Team:Osaka/week3#August 8 (Sun)|8]]<span style={{{s8|"color:{{{c8|red}}};"}}}></span>
 +
|[[Team:Osaka/week3#August 9 (Mon)|9]]<span style={{{s9|"color:{{{c9|inherit}}};"}}}></span>
 +
|[[Team:Osaka/week3#August 10 (Tue)|10]]<span style={{{s10|"color:{{{c10|inherit}}};"}}}></span>
 +
|[[Team:Osaka/week3#August 11 (Wed)|11]]<span style={{{s11|"color:{{{c11|inherit}}};"}}}></span>
 +
|<span style={{{s12|"color:{{{c12|inherit}}};"}}}>12</span>
 +
|<span style={{{s13|"color:{{{c13|inherit}}};"}}}>13</span>
 +
|<span style={{{s14|"color:{{{c14|deepskyblue}}};"}}}>14</span>
|-
|-
-
|F2||N/A||C||RBS + F1
+
|<span style>[[Team:Osaka/week4|week4]]</span>
 +
|<span style={{{s15|"color:{{{c15|red}}};"}}}>15</span>
 +
|[[Team:Osaka/week4#August 16 (Mon)|16]]<span style={{{s16|"color:{{{c16|inherit}}};"}}}></span>
 +
|[[Team:Osaka/week4#August 17 (Tue)|17]]<span style={{{s17|"color:{{{c17|inherit}}};"}}}></span>
 +
|[[Team:Osaka/week4#August 18 (Wed)|18]]<span style={{{s18|"color:{{{c18|inherit}}};"}}}></span>
 +
|[[Team:Osaka/week4#August 19 (Thu)|19]]<span style={{{s19|"color:{{{c19|inherit}}};"}}}></span>
 +
|[[Team:Osaka/week4#August 20 (Fri)|20]]<span style={{{s20|"color:{{{c20|inherit}}};"}}}></span>
 +
|[[Team:Osaka/week4#August 21 (Sat)|21]]<span style={{{s21|"color:{{{c21|deepskyblue}}};"}}}></span>
|-
|-
-
|F3||N/A||C||Lac promoter + RBS + F1
+
|<span style>[[Team:Osaka/week5|week5]]</span>
-
|}
+
|[[Team:Osaka/week5#August 22 (Sun)|22]]<span style={{{s22|"color:{{{c22|red}}};"}}}></span>
-
 
+
|[[Team:Osaka/week5#August 23 (Mon)|23]]<span style={{{s23|"color:{{{c23|inherit}}};"}}}></span>
-
===August 23 (Mon)===
+
|[[Team:Osaka/week5#August 24 (Tue)|24]]<span style={{{s24|"color:{{{c24|inherit}}};"}}}></span>
-
1.液培へ
+
|[[Team:Osaka/week5#August 25 (Wed)|25]]<span style={{{s25|"color:{{{c25|inherit}}};"}}}></span>
-
    (1),(2),(3),2-4A
+
|[[Team:Osaka/week5#August 26 (Thu)|26]]<span style={{{s26|"color:{{{c26|inherit}}};"}}}></span>
-
    LB 3ml,Amp 3μl,Chl 1μl                ←濃度で表したほうがいいのでは？？
+
|[[Team:Osaka/week5#August 27 (Fri)|27]]<span style={{{s27|"color:{{{c27|inherit}}};"}}}></span>
-
    incubate 37℃ 9:30~
+
|[[Team:Osaka/week5#August 28 (Sat)|28]]<span style={{{s28|"color:{{{c28|deepskyblue}}};"}}}></span>
-
 
+
|-
-
2.mini prep プラスミド回収
+
|<span style>[[Team:Osaka/week6|week6]]</span>
-
    1-1D・1-2M(3)
+
|[[Team:Osaka/week6#August 29 (Sun)|29]]<span style={{{s29|"color:{{{c29|red}}};"}}}></span>
-
 
+
|[[Team:Osaka/week6#August 30 (Mon)|30]]<span style={{{s30|"color:{{{c30|inherit}}};"}}}></span>
-
3.制限酵素処理
+
|[[Team:Osaka/week6#August 31 (Tue)|31]]<span style={{{s31|"color:{{{c31|inherit}}};"}}}></span>
-
    プラスミド 1-1D・1-2M(3)    2.5μl
+
|colspan="4"|
-
    EcoRI            1μl
+
|- style="background:{{{color3|#eeeeff}}};"
-
    SpeI            1μl
+
|colspan="7"|{{{footer|}}}
-
    H2O            40μl
+
|}<noinclude>
-
    10xNEBuffer2        5μl
+
{| class="toccolours" style="float:{{{float|left}}}; width:{{{width|240px}}}; text-align:center;"
-
    100xBSA            0.5μl
+
|- style="background:{{{color1|#ccccff}}};"
-
    total            50μl
+
|colspan="8"|{{{header|September}}}
-
 
+
|-
-
    37℃ incubate 10:30~
+
|- style="background:{{{color2|#eeeeff}}};"
-
 
+
|width=100px|Week
-
　電気泳動
+
|width="14%"|<span style="color:red;">S</span>
-
    dye 2μl,サンプル 10μl(ネガコン 2μl),Ladder 2μl    1%Agalose gel,100V 20min,EtBr 30min
+
|width="14%"|M
-
    Ladder,1-1D・1-2M d,1-1D・1-2M
+
|width="14%"|T
-
 
+
|width="14%"|W
-
4.液培へ
+
|width="14%"|T
-
    1-1D・1-2M    3つ
+
|width="14%"|F
-
    12:53~ 37℃ incubate
+
|width="14%"|<span style="color:deepskyblue;">S</span>
-
 
+
|-
-
5.トランスフォーメーション
+
|<span style>[[Team:Osaka/week6|week6]]</span>
-
    2-2O,3-11I    Amp
+
|<span style></span>
-
    13:30~ 37℃
+
|<span style></span>
-
 
+
|<span style></span>
-
===August 24 (Tue)===
+
|[[Team:Osaka/week6#September 1 (Wed)|1]]<span style={{{s1|"color:{{{c1|inherit}}};"}}}></span>
-
1.transformation結果
+
|[[Team:Osaka/week6#September 2 (Thu)|2]]<span style={{{s2|"color:{{{c2|inherit}}};"}}}></span>
-
    2-2O    100以上
+
|[[Team:Osaka/week6#September 3 (Fri)|3]]<span style={{{s3|"color:{{{c3|inherit}}};"}}}></span>
-
    3-11I    100以上
+
|[[Team:Osaka/week6#September 4 (Sat)|4]]<span style={{{s4|"color:{{{c4|deepskyblue}}};"}}}></span>
-
            →液培に
+
|-
-
 
+
|<span style>[[Team:Osaka/week7|week7]]</span>
-
2.miniprep プラスミド回収
+
|[[Team:Osaka/week7#September 5 (Sun)|5]]<span style={{{s5|"color:{{{c5|red}}};"}}}></span>
-
    F1,F2,F3,2-4A    (8/22にトラフォした(1)~(3)はF1~3に名前を変更しました。)
+
|[[Team:Osaka/week7#September 6 (Mon)|6]]<span style={{{s6|"color:{{{c6|inherit}}};"}}}></span>
-
 
+
|[[Team:Osaka/week7#September 7 (Tue)|7]]<span style={{{s7|"color:{{{c7|inherit}}};"}}}></span>
-
3.制限処理酵素
+
|[[Team:Osaka/week7#September 8 (Wed)|8]]<span style={{{s8|"color:{{{c8|inherit}}};"}}}></span>
-
    プラスミド F1,F2,F3        2.5μl
+
|[[Team:Osaka/week7#September 9 (Thu)|9]]<span style={{{s9|"color:{{{c9|inherit}}};"}}}></span>
-
    EcoRI            1μl
+
|[[Team:Osaka/week7#September 10 (Fri)|10]]<span style={{{s10|"color:{{{c10|inherit}}};"}}}></span>
-
    SpeI            1μl
+
|[[Team:Osaka/week7#September 11 (Sat)|11]]<span style={{{s11|"color:{{{c11|deepskyblue}}};"}}}></span>
-
    H2O            40μl
+
|-
-
    10xNEBuffer2        5μl
+
|<span style>[[Team:Osaka/week8|week8]]</span>
-
    100xBSA            0.5μl
+
|[[Team:Osaka/week8#September 12 (Sun)|12]]<span style={{{s12|"color:{{{c12|red}}};"}}}></span>
-
    total            50μl
+
|[[Team:Osaka/week8#September 13 (Mon)|13]]<span style={{{s13|"color:{{{c13|inherit}}};"}}}></span>
-
 
+
|[[Team:Osaka/week8#September 14 (Tue)|14]]<span style={{{s14|"color:{{{c14|inherit}}};"}}}></span>
-
    37℃ incubate 11:30~
+
|[[Team:Osaka/week8#September 15 (Wed)|15]]<span style={{{s15|"color:{{{c15|inherit}}};"}}}></span>
-
 
+
|[[Team:Osaka/week8#September 16 (Thu)|16]]<span style={{{s16|"color:{{{c16|inherit}}};"}}}></span>
-
    F3をもう一つ行う→F3'とする
+
|[[Team:Osaka/week8#September 17 (Fri)|17]]<span style={{{s17|"color:{{{c17|inherit}}};"}}}></span>
-
    プラスミド 1F        2.5μl
+
|[[Team:Osaka/week8#September 18 (Sat)|18]]<span style={{{s18|"color:{{{c18|deepskyblue}}};"}}}></span>
-
    EcoRI            1μl
+
|-
-
    PstI            1μl
+
|<span style>[[Team:Osaka/week9|week9]]</span>
-
    H2O            40μl
+
|[[Team:Osaka/week9#September 19 (Sun)|19]]<span style={{{s19|"color:{{{c19|red}}};"}}}></span>
-
    10xNEBuffer2        5μl
+
|[[Team:Osaka/week9#September 20 (Mon)|20]]<span style={{{s20|"color:{{{c20|inherit}}};"}}}></span>
-
    100xBSA            0.5μl
+
|[[Team:Osaka/week9#September 21 (Tue)|21]]<span style={{{s21|"color:{{{c21|inherit}}};"}}}></span>
-
    total            50μl
+
|[[Team:Osaka/week9#September 22 (Wed)|22]]<span style={{{s22|"color:{{{c22|inherit}}};"}}}></span>
-
 
+
|[[Team:Osaka/week9#September 23 (Thu)|23]]<span style={{{s23|"color:{{{c23|inherit}}};"}}}></span>
-
    37℃ incubate 12:53~
+
|[[Team:Osaka/week9#September 24 (Fri)|24]]<span style={{{s24|"color:{{{c24|inherit}}};"}}}></span>
-
 
+
|[[Team:Osaka/week9#September 25 (Sat)|25]]<span style={{{s25|"color:{{{c25|deepskyblue}}};"}}}></span>
-
4.電気泳動
+
|-
-
    dye 2μl,サンプル 10μl,Ladder 2μl    1% Agalose gel,100V 30min,EtBr 30min
+
|<span style>[[Team:Osaka/week10|week10]]</span>
-
    Ladder,F1 d,F2 d,F3 d
+
|[[Team:Osaka/week10#September 26 (Sun)|26]]<span style={{{s26|"color:{{{c26|red}}};"}}}></span>
-
 
+
|[[Team:Osaka/week10#September 27 (Mon)|27]]<span style={{{s27|"color:{{{c27|inherit}}};"}}}></span>
-
    dye 2μl,サンプル 10μl(ネガコン 2μl),Ladder 2μl    1% Agalose gel,100V 30min,EtBr 30min
+
|[[Team:Osaka/week10#September 28 (Tue)|28]]<span style={{{s28|"color:{{{c28|inherit}}};"}}}></span>
-
    F3,F3' d,Ladder
+
|[[Team:Osaka/week10#September 29 (Wed)|29]]<span style={{{s29|"color:{{{c29|inherit}}};"}}}></span>
-
 
+
|[[Team:Osaka/week10#September 30 (Thu)|30]]<span style={{{s30|"color:{{{c30|inherit}}};"}}}></span>
-
5.液培へ
+
|colspan="2"|
-
    F1    LB 3ml,Amp 3μ
+
|- style="background:{{{color3|#eeeeff}}};"
-
    37℃ incubate 14:30~
+
|colspan="7"|{{{footer|}}}
-
 
+
|}<noinclude>
-
6.グリセロールストック作り
+
{| class="toccolours" style="float:{{{float|left}}}; width:{{{width|240px}}}; text-align:center;"
-
    (1-1D,1-2M,1-3A)の培養液 200mlと50%グリセリン 100mlを混合し-80℃へ
+
|- style="background:{{{color1|#ccccff}}};"
-
 
+
|colspan="8"|{{{header|October}}}
-
===August 25 (Wed)===
+
|-
-
1.miniprep プラスミド回収
+
|- style="background:{{{color2|#eeeeff}}};"
-
    F1,3-11I,2-2O
+
|width=100px|Week
-
 
+
|width="14%"|<span style="color:red;">S</span>
-
2.制限酵素処理
+
|width="14%"|M
-
    プラスミド F1,3-11I        2.5μl
+
|width="14%"|T
-
    EcoRI            1μl
+
|width="14%"|W
-
    SpeI            1μl
+
|width="14%"|T
-
    H2O            40μl
+
|width="14%"|F
-
    10xNEBuffer2        5μl
+
|width="14%"|<span style="color:deepskyblue;">S</span>
-
    100xBSA            0.5μl
+
|-
-
    total            50μl
+
|<span style>[[Team:Osaka/week10|week10]]</span>
-
 
+
|<span style></span>
-
    37℃ incubate 10:45~
+
|<span style></span>
-
 
+
|<span style></span>
-
3.電気泳動
+
|<span style></span>
-
    dye 2μl,サンプル 10μl,Ladder 2μl    1% Agalose gel,100V 30min,EtBr 40min
+
|<span style></span>
-
    Ladder,3-11I d,F1 d
+
|[[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>[[Team:Osaka/week11|week11]]</span>
 +
|[[Team:Osaka/week11#October 3 (Sun)|3]]<span style={{{s3|"color:{{{c3|red}}};"}}}></span>
 +
|[[Team:Osaka/week11#October 4 (Mon)|4]]<span style={{{s4|"color:{{{c4|inherit}}};"}}}></span>
 +
|[[Team:Osaka/week11#October 5 (Tue)|5]]<span style={{{s5|"color:{{{c5|inherit}}};"}}}></span>
 +
|[[Team:Osaka/week11#October 6 (Wed)|6]]<span style={{{s6|"color:{{{c6|inherit}}};"}}}></span>
 +
|[[Team:Osaka/week11#October 7 (Thu)|7]]<span style={{{s7|"color:{{{c7|inherit}}};"}}}></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>[[Team:Osaka/week12|week12]]</span>
 +
|[[Team:Osaka/week12#October 10 (Sun)|10]]<span style={{{s10|"color:{{{c10|red}}};"}}}></span>
 +
|[[Team:Osaka/week12#October 11 (Mon)|11]]<span style={{{s11|"color:{{{c11|inherit}}};"}}}></span>
 +
|[[Team:Osaka/week12#October 12 (Tue)|12]]<span style={{{s12|"color:{{{c12|inherit}}};"}}}></span>
 +
|[[Team:Osaka/week12#October 13 (Wed)|13]]<span style={{{s13|"color:{{{c13|inherit}}};"}}}></span>
 +
|[[Team:Osaka/week12#October 14 (Thu)|14]]<span style={{{s14|"color:{{{c14|inherit}}};"}}}></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>[[Team:Osaka/week13|week13]]</span>
 +
|[[Team:Osaka/week13#October 17 (Sun)|17]]<span style={{{s17|"color:{{{c17|red}}};"}}}></span>
 +
|[[Team:Osaka/week13#October 18 (Mon)|18]]<span style={{{s18|"color:{{{c18|inherit}}};"}}}></span>
 +
|[[Team:Osaka/week13#October 19 (Tue)|19]]<span style={{{s19|"color:{{{c19|inherit}}};"}}}></span>
 +
|[[Team:Osaka/week13#October 20 (WEd)|20]]<span style={{{s20|"color:{{{c20|inherit}}};"}}}></span>
 +
|[[Team:Osaka/week13#October 21 (Thu)|21]]<span style={{{s21|"color:{{{c21|inherit}}};"}}}></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>[[Team:Osaka/week14|week14]]</span>
 +
|[[Team:Osaka/week14#October 24 (Sun)|24]]<span style={{{s24|"color:{{{c24|red}}};"}}}></span>
 +
|[[Team:Osaka/week14#October 25 (Mon)|25]]<span style={{{s25|"color:{{{c25|inherit}}};"}}}></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={{{s29|"color:{{{c29|inherit}}};"}}}>29</span>
 +
|<span style={{{s30|"color:{{{c30|deepskyblue}}};"}}}>30</span>
 +
|-
 +
|<span style>[[Team:Osaka/week15|week15]]</span>
 +
|[[{{{prefix|}}}10.31{{{suffix|}}}|<span style={{{s31|"color:{{{c31|red}}};"}}}>31</span>]]
 +
|colspan="6"|
 +
|- style="background:{{{color3|#eeeeff}}};"
 +
|colspan="7"|{{{footer|}}}
 +
|}<noinclude>
Line 294: Line 361:
<html>
<html>
</div>
</div>
 +
</div>
 +
</div>
 +
<div class="clear"><hr></div>
 +
</html>
 +
{{Osaka_foot}}
 +
<html>
 +
</html>
</html>

Latest revision as of 03:34, 28 October 2010

Notebook

July / August
Week S M T W T F S
week1 25 26 27 28 29 30 31
week2 1 2 3 4 5 6 7
week3 8 9 10 11 12 13 14
week4 15 16 17 18 19 20 21
week5 22 23 24 25 26 27 28
week6 29 30 31
September
Week S M T W T F S
week6 1 2 3 4
week7 5 6 7 8 9 10 11
week8 12 13 14 15 16 17 18
week9 19 20 21 22 23 24 25
week10 26 27 28 29 30
October
Week S M T W T F S
week10 1 2
week11 3 4 5 6 7 8 9
week12 10 11 12 13 14 15 16
week13 17 18 19 20 21 22 23
week14 24 25 26 27 28 29 30
week15 31



© iGEM OSAKA 2010 All rights reserved

Retrieved from "http://2010.igem.org/Team:Osaka/Notebook"