User:Jorge Arturo Zepeda/Notebook/iGem LCG-UNAM team 2010/2010/04/21
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Wednesday 21st April 2010
I cultivated the 5 colonies resulting from the Monday transformation in 4ml of liquid lb with 4ul of kanamycin and also a medium control at 9 a.m.
I came back a 3 p.m. to extract the plasmid BBa_I712019 (luciferase from Photinus pyralis) with the kit from Roche.
For this I centrifuged 3 times the LB to get the whole pellet of bacteria from each tube, alter that I confused the plasmid extraction kit with the PCR exctraction kit, and I resuspended the pellet in 80ul of water, but then I realizad my mistake and centrifugues again to get the pellet, afterwards I extracted the plasmid from each sample using the Roche kit.
Double restriction with EcoR1 and PST:
I prepared a stock for the 5 reactions using the calculations for 6 as it follows:
Reactive....1x [ul]......6x [ul]