Team:Chiba/Calendar-Home/13 September 2008

From 2008.igem.org

(Difference between revisions)
m (→Team:Input)
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Time Delay Test
Time Delay Test
-
①T9002(Ptet) -p15a-Kan(BW)
+
#T9002(Ptet) -p15a-Kan(BW)
 +
#Plac-RhlI -Amp(BW)
 +
#Plac-LasI -Amp(BW)
 +
#Ptet-LuxI -Amp(BW)
-
②Plac-RhlI -Amp(BW)
+
*pre-culture
 +
#LB-Kan 2ml培養
-
③Plac-LasI -Amp(BW)
+
2,3:LB-Amp -0.2%グルコース 2ml培養
-
④Ptet-LuxI -Amp(BW)
+
4LB-Amp　2ml培養
-
 
+
-
*プレ培
+
-
①LB-Kan 2ml培養
+
-
 
+
-
②、③LB-Amp -0.2%グルコース 2ml培養
+
-
 
+
-
④LB-Amp　2ml培養
+
↓over night
↓over night
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-
48欠Deep Well
+
48-well plate(Deep Well)
<table width="300" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
<table width="300" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
-
<td width="257">①:2ml</td>
+
<td width="257">1:2ml</td>
-
<td>②:2ml,AHL</td><td>②:2ml</td><td>③:2ml</td><td>④2ml</td><td>-</td>
+
<td>2:2ml,AHL</td><td>2:2ml</td><td>3:2ml</td><td>4:2ml</td><td>-</td>
<tr>
<tr>
<td>-</td>
<td>-</td>
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</tr>
</tr>
<tr>
<tr>
-
<td>①:1ml,②:1ml,IPTG:2μl</td>
+
<td>1:1ml,2:1ml,IPTG:2μl</td>
-
<td>①:100μl,②:1ml,IPTG:1.1μl</td><td>①10μl,②1ml,IPTG:101μl</td><td>①:1μl,②1ml,IPTG:1.001μl</td><td>①:1ml,②:100μl,IPTG:1.1μl</td><td>①:1ml,②:10μl,IPTG:1.01μl</td>
+
<td>1:100&mu;l,2:1ml,IPTG:1.1&mu;l</td><td>110&mu;l,21ml,IPTG:101&mu;l</td><td>1:1μl,21ml,IPTG:1.001&mu;l</td><td>1:1ml,2:100&mu;l,IPTG:1.1&mu;l</td><td>1:1ml,2:10&mu;l,IPTG:1.01&mu;l</td>
</tr>
</tr>
<tr>
<tr>
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</tr>
</tr>
<tr>
<tr>
-
<td>①:1ml,③:1ml,IPTG:2μl</td>
+
<td>1:1ml,3:1ml,IPTG:2&mu;l</td>
-
<td>①:100μl,③:1ml,IPTG:1.1μl</td><td>①:10μl,③:1ml,IPTG:1.01μl</td><td>①:1μl,③:1ml,IPTG</td><td>①:1ml,③:100μl,IPTG:1.1μl</td><td>①:1ml,③:10μl,IPTG:1.01μl</td>
+
<td>1:100&mu;l,3:1ml,IPTG:1.1&mu;l</td><td>1:10&mu;l,3:1ml,IPTG:1.01&mu;l</td><td>1:1μl,3:1ml,IPTG</td><td>1:1ml,3:100&mu;l,IPTG:1.1&mu;l</td><td>1:1ml,3:10&mu;l,IPTG:1.01&mu;l</td>
</tr>
</tr>
<tr>
<tr>
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</tr>
</tr>
<tr>
<tr>
-
<td>①:1ml,④:1ml,IPTG:2μl</td>
+
<td>1:1ml,4:1ml,IPTG:2&mu;l</td>
-
<td>①100μl,④:1ml,IPTG:1.1μl</td><td>①:10μl,④:1ml,IPTG:1.0μl</td><td>①:1μl,④:1ml,IPTG:1.001μl</td><td>①:1ml,④:100μl,IPTG:1.0μl</td><td>①:1ml,④:10μl,IPTG1.01μl</td>
+
<td>1:100&mu;l,4:1ml,IPTG:1.1&mu;l</td><td>1:10&mu;l,4:1ml,IPTG:1.0&mu;l</td><td>1:1&mu;l,4:1ml,IPTG:1.001&mu;l</td><td>1:1ml,4:100&mu;l,IPTG:1.0&mu;l</td><td>1:1ml,4:10&mu;l,IPTG1.01&mu;l</td>
</tr>
</tr>
<tr>
<tr>

Revision as of 19:40, 29 October 2008

>Home | Notebook

12 September 2008 <|> 14 September 2008

Contents

Laboratory work

Team:Input

Time Delay Test

  1. T9002(Ptet) -p15a-Kan(BW)
  2. Plac-RhlI -Amp(BW)
  3. Plac-LasI -Amp(BW)
  4. Ptet-LuxI -Amp(BW)
  • pre-culture
  1. LB-Kan 2ml培養

2,3:LB-Amp -0.2%グルコース 2ml培養

4LB-Amp　2ml培養

↓over night

  • 本培

50ml遠心管に①の菌液100μlとLB-Kanを20mlを加える。

50ml遠心管に②、③それぞれの菌液10μlとLB-Amp 0.2%グルコースをそれぞれに6mlを加える。

50ml遠心管に④のLB-Ampを6mlを加える。

OD>2となったらWash


  • ①に関して

①

↓冷却遠心(3700rpm,3min)　　　　　　　　　　

上澄みを捨てる

LB-Kanを20ml加える。　　　　　　　　　　　　


  • ②③④に関して

②、③、④

↓冷却遠心(3700rpm,3min)

上澄みを捨てる。

LBを2ml加える。　　　　　　　　　　　　

↓冷却遠心(3700rpm,3min)

上澄みを捨てる。

LBを2ml加える。

↓冷却遠心(3700rom,3min)

上澄み捨てる。

LB-Ampを6ml加える。


48-well plate(Deep Well)

1:2ml 2:2ml,AHL2:2ml3:2ml4:2ml-
- -----
1:1ml,2:1ml,IPTG:2μl 1:100μl,2:1ml,IPTG:1.1μl110μl,21ml,IPTG:101μl1:1μl,21ml,IPTG:1.001μl1:1ml,2:100μl,IPTG:1.1μl1:1ml,2:10μl,IPTG:1.01μl
- -----
1:1ml,3:1ml,IPTG:2μl 1:100μl,3:1ml,IPTG:1.1μl1:10μl,3:1ml,IPTG:1.01μl1:1μl,3:1ml,IPTG1:1ml,3:100μl,IPTG:1.1μl1:1ml,3:10μl,IPTG:1.01μl
- -----
1:1ml,4:1ml,IPTG:2μl 1:100μl,4:1ml,IPTG:1.1μl1:10μl,4:1ml,IPTG:1.0μl1:1μl,4:1ml,IPTG:1.001μl1:1ml,4:100μl,IPTG:1.0μl1:1ml,4:10μl,IPTG1.01μl
- -----

Team:Communication

Gel Check
  1. [http://partsregistry.org/Part:BBa_Co161 BBa_C0161](2007)
  2. [http://partsregistry.org/Part:BBa_J04500 BBa_J04500](2007)
  3. [http://partsregistry.org/Part:BBa_R0010 BBa_R0010](2007)
Chiba-0913.JPG
Sample No. 1~3
Sample DNA(μL) 1
Loading Dye(μL) 1
dH2O(μL) 4
TOTAL(μL) 6
From left;


-->Digestion

  1. [http://partsregistry.org/Part:BBa_Co161 BBa_C0161](2007)
  2. [http://partsregistry.org/Part:BBa_J04500 BBa_J04500](2007)
  3. [http://partsregistry.org/Part:BBa_R0010 BBa_R0010](2007)


Sample No. ①②③④
Sample DNA(μL) 5511
EcoRⅠ(μL) 0.5o.5--
SpeⅠ(μL) 0.50.5--
Buffer 1(μL) 11--
Buffer 3(μL) --1-
BSA(μL) 11--
dH2O(μL) 227.5-
TOTAL(μL) 1010101

Team:Output

no work

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