IGEM:Paris Bettencourt 2012/Notebooks/Delay group/day by day//2012/07/26: Difference between revisions
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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 | ||
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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 09:56, 26 July 2012
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Recovery1- 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. Digestioninsert = 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 |