Haynes Lab:Notebook/CRISPR Editing/2015/09/10

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09/10/2015

Ran g031 P197/198 PCR samples on gel. No band in water-only control PCR. Gal4-EED +dox g031 rep 2 failed. Re-ran with 5ul of gDNA and it worked.
Nested PCR of g031 with P149/P174. No band in water-only control PCR.

gDNA extraction of g025 and g044 treated cells

Cells drug gRNA rep 260 280 260/280 ng/µL ul for 200 ul water 15-
Gal4-EED puro g025 1 0.015 0.008 1.973 15.45 12.94 2.06
Gal4-EED puro g025 2 0.029 0.015 1.993 28.969 6.90 8.10
Gal4-EED puro g025 3 0.02 0.01 2.032 20.101 9.95 5.05
Gal4-EED dox g025 1 0.037 0.019 2.006 37.202 5.38 9.62
Gal4-EED dox g025 2 0.08 0.041 1.974 80.301 2.49 12.51
Gal4-EED dox g025 3 0.075 0.039 1.926 74.544 2.68 12.32
Gal4-EED puro g044 1 0.015 0.01 1.522 14.529 13.77 1.23
Gal4-EED puro g044 2 0.023 0.012 1.893 23.315 8.58 6.42
Gal4-EED puro g044 3 0.05 0.025 2.008 49.979 4.00 11.00
Gal4-EED dox g044 1 0.034 0.016 2.102 33.619 5.95 9.05
Gal4-EED dox g044 2 0.069 0.037 1.881 69.057 2.90 12.10
Gal4-EED dox g044 3 0.044 0.023 1.917 44.009 4.54 10.46

Set up phusion PCR with P197/198 with 200ng gDNA