Haynes Lab:Notebook/CRISPR Editing/2016/05/24

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05/24/2015

Added RNase to g025 sonicated chromatin, 30min at 37

finally got enough DNA from PCR for g025 3. Means there's probably something wrong with my Phusion.

Cell type gRNA rep 260 280 260/280 ng/µL ul for 400ng ul hyb buff ul H2O
Luc14g02530.0340.0191.80933.88511.801.501.70

Can run surveyor on g025 and 31 today. Set up annealing reaction:

Cell type gRNA rep 260 280 260/280 ng/µL ul for 400ng ul hyb buff ul water
Luc142510.0350.021.79235.27911.341.502.16
Luc142520.0460.0251.87646.2548.651.504.85
Luc142530.0340.0191.80933.88511.801.501.70
Luc143110.070.0371.969.9825.721.507.78
Luc143120.0870.0461.88586.9154.601.508.90
Luc143130.0350.0191.86835.48411.271.502.23



Added proteinase k to 25 sonicated chromatin. 62 for 2 hours

conc of sheared chromatin

Cells gRNA rep 260 280 260/280 ng/µL ul for 50ug ul dil buff
Luc142512.2721.321.7212271.86422.01177.99
Luc142522.4261.421.7082426.16520.61179.39
Luc142532.0221.171.7282021.66424.73175.27
Gal42512.1281.2671.682127.68823.50176.50
Gal42521.3050.7951.6431305.14838.31161.69
Gal42531.9031.1381.6721902.94126.28173.72



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