Haynes Lab:Notebook/CRISPR Editing/2015/08/31

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08/31/2015

Surveyor assay on Luc14 dilutions in triplicate, 1ug, 0.1ug, and 0.01ug g034. 1ul surveyor, 1ul enhancer, 1hour at 42deg. after 1h, added 1.9ul of stop solution, diluted 1:20 in water, ran on BA.

Surveyor assay on Luc14 singlet failed gRNAs. 1ul surveyor, 1ul enhancer, 1hour at 42deg. after 1h, added 1.9ul of stop solution, diluted 1:20 in water, ran on BA.



Phusion PCR for sequencing luciferase. template: sheared, untreated, Luc14 gDNA

F primer R primer exp size Tann seq primer
P218 P233 1087 61 P218, 283
P161 P233 1799 61 P175
P169 P233 1249 61 P171


PCR purified the samples from the dilution experiment that didn't have enough DNA.

Taq PCR P166/238 sheared untreated Luc14 gDNA, at 47, ext 2:30, length, 2259bp