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

(Difference between revisions)

Revision as of 12:56, 26 July 2012

Project name

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.

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 :

Digestion of the vector :

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

@@ Line 20: / Line 20: @@
 - pLac-RBS-GFP-TT clone 1 (From Denis) + Amp
-- pBad-RBS-LacI clone 2 (from the 2009 Paris iGEM team) + Amp
+- araC-pBad-RBS-LacI clone 2 (from the 2009 Paris iGEM team) + Amp
-- pBad-RBS-LacI clone 2 (from the 2009 Paris iGEM team) + Kan
+- araC-pBad-RBS-LacI clone 2 (from the 2009 Paris iGEM team) + Kan
 - pLac-RBS-GFP-TT clone 2 (From Antoine) +IPTG+Amp
@@ Line 31: / Line 31: @@
 - 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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