Haynes Lab:Notebook/CRISPR Editing/2015/03/22

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03/22/2015

Column purified unt and g032 treated nested PCR with P149/174. Will set up hybridization of 400ng PCR product in 15ul reaction:

Cell type treatment rep 260 280 260/280 ng/┬ÁL ul for 400ng ul water ul hyb buff total
Luc14Unt10.0360.021.80536.0911.082.421.515
Luc14Unt20.0420.0241.80142.3499.454.051.515
Luc14Unt30.0380.0211.81538.21910.473.031.515
Gal4-EED+doxUnt10.0450.0251.77745.018.894.611.515
Gal4-EED+doxUnt20.0420.0231.83642.289.464.041.515
Gal4-EED+doxUnt30.0380.0221.7738.20110.473.031.515
Luc14g03210.0350.0181.98235.03511.422.081.515
Luc14g03220.040.0221.78739.67610.083.421.515
Luc14g03230.0420.0241.78542.1769.484.021.515
Gal4-EED+doxg03210.0430.0231.85142.7659.354.151.515
Gal4-EED+doxg03220.0370.0221.6536.65810.912.591.515
Gal4-EED+doxg03230.0380.0221.74237.77510.592.911.515


Annealing reaction in Surveyor protocol
1ul surveyor enzyme, 1ul enhancer, 60 mins at 42deg
Dilute 1:10 in ES, run on bioanalyzer



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