Haynes Lab:Notebook/CRISPR Editing/2015/03/12

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03/12/2015

Sample Read# mL culture elution 260 280 260/280 ng/µL
KAH228 2 1 0.12 0.064 1.885 120.452
KAH228 2 2 0.019 0.009 2.056 18.943
KAH228 6 1 0.383 0.205 1.87 382.939
KAH228 6 2 0.038 0.02 1.894 38.18
KAH228 12 1 0.376 0.201 1.873 376.089
KAH228 12 2 0.044 0.025 1.762 43.806



Cut 250ng of g031 treated luc14 and gal4eed+dox P151/158 nested PCR products. one hour at 30degC then heat inactivated at 65 for 20 minutes. No enzyme control has 13ul water, 0ul enzyme:

water 12.5
DNA 5
Buffer 2
Bsa1I 0.5
total 20

Ran on BA, saw lots of uncut amplicons, equal for Luc14 and Gal4, think the enzyme didn't cut completely.