IGEM:Paris Bettencourt 2012/Notebooks/Semantic group/day by day//2012/08/29: Difference between revisions
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for the control, I strictly follow the protocol. | for the control, I strictly follow the protocol. | ||
After PCR, I let the PCR tube on ice for 5 min, then I add 1µL of DpnI, and transfered in 1.5 mL eppendorf tube. The digestion proceed in waterbath at 37°C for 1h. | |||
I transformed then 1µL of the digestion product into 50µL of XL-1 super competent cells. | |||
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Revision as of 09:58, 29 August 2012
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Quick Change Mutagenesislink of the protocol : QCM protocol First I dilute the primers I received in 77 µL and 139 µL for the Fw and Rev respectively, that give the primers at a concentration of 100µM. Then I diluted 10 times to get aliquots of 10µM of primers, in 50µL. I use 2µL of this aliquot for the PCR. I have 3 samples, with respectively 17, 34 and 51 ng of dna for the PCR. The PCR program is :
for the control, I strictly follow the protocol. After PCR, I let the PCR tube on ice for 5 min, then I add 1µL of DpnI, and transfered in 1.5 mL eppendorf tube. The digestion proceed in waterbath at 37°C for 1h. I transformed then 1µL of the digestion product into 50µL of XL-1 super competent cells. | |
