IGEM:Paris Bettencourt 2012/Notebooks/Delay group/day by day//2012/07/26

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(Entry title)
(Recovery)
Line 20: Line 20:
6- pLac-RBS-GFP-TT clone 1 (From Denis) + Amp
6- pLac-RBS-GFP-TT clone 1 (From Denis) + Amp
-
7- pBad-RBS-LacI clone 2 (from the 2009 Paris iGEM team) + Amp
+
7- araC-pBad-RBS-LacI clone 2 (from the 2009 Paris iGEM team) + Amp
-
8- pBad-RBS-LacI clone 2 (from the 2009 Paris iGEM team) + Kan
+
8- araC-pBad-RBS-LacI clone 2 (from the 2009 Paris iGEM team) + Kan
9- pLac-RBS-GFP-TT clone 2 (From Antoine) +IPTG+Amp
9- pLac-RBS-GFP-TT clone 2 (From Antoine) +IPTG+Amp
Line 31: Line 31:
12- pLac-RBS-GFP-TT clone 4 (From Antoine) +Amp
12- pLac-RBS-GFP-TT clone 4 (From Antoine) +Amp
 +
 +
We'll need to do a glycerol stock for the strains that worked.
 +
 +
==Digestion==
 +
 +
insert = pLac-GFP-TT
 +
 +
vector = araC-pBad-RBS-LacI
 +
 +
The insert is digested in E&S, the vector is digested in E&X.
 +
 +
Digestion of the insert :
 +
*13µl insert (150 ng/µl)
 +
*2µl EcoRI
 +
*2µl SpeI
 +
*2µl FD Green Buffer
 +
*1µl H2O
 +
 +
Digestion of the vector :
 +
*11µl vector (180 ng/µl)
 +
*2µl EcoRI
 +
*2µl XbaI
 +
*2µl FD Buffer
 +
*3µl H2O
 +
 +
The insert is purified on a 0.8% agarose gel. We obtain a concentration of 24.1 ng/µl
 +
[[Image:KR003886.png|center|500px]]
 +
The vector is purified wth the PCR kit. We obtain a concentration of 86.4 ng/µl
 +
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Revision as of 12:56, 26 July 2012

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Recovery

1- pLac-RBS-GFP-TT clone 1 (From Antoine) +IPTG+Amp

2- pLac-RBS-GFP-TT clone 1 (From Antoine) +Amp

3- pLac-RBS-GFP-TT clone 1 (From Antoine) +IPTG+Kan

4- pLac-RBS-GFP-TT clone 1 (From Antoine) +Kan

5- pLac-RBS-GFP-TT clone 1 (From Denis) + IPTG + Amp

6- pLac-RBS-GFP-TT clone 1 (From Denis) + Amp

7- araC-pBad-RBS-LacI clone 2 (from the 2009 Paris iGEM team) + Amp

8- araC-pBad-RBS-LacI clone 2 (from the 2009 Paris iGEM team) + Kan

9- pLac-RBS-GFP-TT clone 2 (From Antoine) +IPTG+Amp

10- pLac-RBS-GFP-TT clone 2 (From Antoine) +Amp

11- pLac-RBS-GFP-TT clone 4 (From Antoine) +IPTG+Amp

12- pLac-RBS-GFP-TT clone 4 (From Antoine) +Amp

We'll need to do a glycerol stock for the strains that worked.

Digestion

insert = pLac-GFP-TT

vector = araC-pBad-RBS-LacI

The insert is digested in E&S, the vector is digested in E&X.

Digestion of the insert :

  • 13µl insert (150 ng/µl)
  • 2µl EcoRI
  • 2µl SpeI
  • 2µl FD Green Buffer
  • 1µl H2O

Digestion of the vector :

  • 11µl vector (180 ng/µl)
  • 2µl EcoRI
  • 2µl XbaI
  • 2µl FD Buffer
  • 3µl H2O

The insert is purified on a 0.8% agarose gel. We obtain a concentration of 24.1 ng/µl

The vector is purified wth the PCR kit. We obtain a concentration of 86.4 ng/µl

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