Haynes Lab:Notebook/HPK-CFP insertion into Gal4EED/Luc using CRISPR/2016/02/01
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QuikChange transformation results:
QuikChange efficiency is about 68%. Picked 8 colonies from each sample plate and set up liquid cultures and streak plates. Today I'm performing 'colony' PCR on the streak plates and then running them on a gel. For the samples that look good (have product in the right size range), I'll do PCR cleanup and submit for sequencing, as well as perform plasmid preps on the pellets of the liquid cultures.
For EM7:ZeoR_v0120, colonies 1, 2, 3, 4, 5, and 7 have the correct size insert (~700 bp). For MV8, colonies 2, 3, 5, 6, 7, and 8 have the correct size insert (~440 bp). Will perform PCR clean-up on all successful PCR products and send 3 from each type of plasmid out for sequencing by the core lab.
Samples for Sequencing
Sequencing Results (2016-02-03)
Got the sequences back for the MV8 samples, they all look terrible. Going to try miniprepping and submitting plasmids instead.