Haynes Lab:Notebook/CRISPR Editing/2015/01/08

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01/08/2015

Ran the PCR reactions from yesterday on a gel. They look great, no extra bands.
Purified the PCR products using Sigma's PCR clean up kit. Eluted in 50ul elution solution.

Sample Read# 260 280 260/280 ng/µL
Untreated 0.107 0.057 1.862 106.778
23 0.096 0.051 1.889 96.073
25 0.1 0.054 1.846 100.394
29 0.104 0.055 1.889 104.281
30 0.099 0.053 1.862 99.101
32 0.093 0.05 1.867 93.338
33 0.091 0.048 1.875 90.77
34 0.093 0.051 1.845 93.316
36 0.094 0.051 1.854 93.841
39 0.097 0.053 1.847 97.225
43 0.089 0.047 1.901 88.782
44 0.03 0.016 1.889 30.391
45 0.087 0.047 1.861 86.634
46 0.079 0.044 1.814 79.106



Set up annealing reaction for the PCR samples

Sample ul for 400ng ul H2O ul anneal buff
Untreated 3.75 9.75 1.5
23 4.16 9.34 1.5
25 3.98 9.52 1.5
29 3.84 9.66 1.5
30 4.04 9.46 1.5
32 4.29 9.21 1.5
33 4.41 9.09 1.5
34 4.29 9.21 1.5
36 4.26 9.24 1.5
39 4.11 9.39 1.5
43 4.51 8.99 1.5
44 13.16 0.34 1.5
45 4.62 8.88 1.5
46 5.06 8.44 1.5

Put at -20 after they finished
Set up PCR wiht phusion for the remaining gRNA treated samples.