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Team:The University of Alberta/29 July 2008
From 2008.igem.org
The Big Whoops
We found the source of the problems we have been having with using NgoMIV and AgeI: the ER0x and BisDx parts dont have the NgoMIV and AgeI sites in them! Not good! We may have to get them resynthesized! But first we will:
- Check ER01 and ER02 for toxicity to determine if we need to have one or both remade
Today
Chris
- Was going to do colony PCR on the transformants from the weekend but someone put the plates in the incubator instead of the fridge! Now they're overgrown and no good :( So....
- I will do PCR on the original ligation using Vf and Vr to determine whether or not Tryp was ligated into J6. If it was then...
- Retransform the ligation and do colony PCR again tomorrow
- I will do PCR on the original ligation using Vf and Vr to determine whether or not Tryp was ligated into J6. If it was then...
UPDATE: PCR looked bad - could be because using a ligation for PCR might not be the optimal template. I'll religate Tryp into J6 and then retransform the construct.
N.B.: The last few PCRs Ive done have had lots o' bands in the negative (water) controls. I've done PCR on all of the bottles of water in the lab to determine which ones are contaminated. Products will be run on a gel tomorrow.
