IGEM:Paris Bettencourt 2012/Notebooks/Delay group/day by day//2012/07/26: Difference between revisions
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Incubate 1h at 22°C (PCR machine) | Incubate 1h at 22°C (PCR machine) | ||
==transformation== | |||
The product of the ligation is transformed on Amp and Kan plates (3µl of the ligation mix for each). PSC001(Amp) is used as positive control (1µl). I forgot the negative control. | |||
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Revision as of 11:13, 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 ligationinsert/vector~5 In 10µl :
Incubate 1h at 22°C (PCR machine) transformationThe product of the ligation is transformed on Amp and Kan plates (3µl of the ligation mix for each). PSC001(Amp) is used as positive control (1µl). I forgot the negative control. |