Haynes Lab:Notebook/CRISPR Editing/2015/03/04

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03/04/2015

Plated cells for transfections tomorrow. Gal4-EED+dox cells plated at 0.16x10^6 cells/well in 12 well plates. Luc14 cells plated at 0.1x10^6 cells / well in 12 well plates. Two plates for each cell type for a total of 4 12-well plates.

Cell type treatment rep 260 280 260/280 ng/µL
Luc14 g023 1 0.089 0.048 1.862 89.331
Luc14 g023 2 0.087 0.046 1.909 87.359
Luc14 g023 3 0.088 0.048 1.834 88.229
Gal4-EED +dox g023 1 0.091 0.049 1.869 91.083
Gal4-EED +dox g023 2 0.091 0.05 1.833 91.397
Gal4-EED +dox g023 3 0.096 0.052 1.832 95.767
Luc14 g045 1 0.078 0.043 1.808 77.703
Luc14 g045 2 0.029 0.016 1.75 28.731
Luc14 g045 3 0.067 0.036 1.861 66.736
Gal4-EED +dox g045 1 0.085 0.046 1.836 85.215
Gal4-EED +dox g045 2 0.073 0.041 1.788 73.213
Gal4-EED +dox g045 3 0.067 0.037 1.805 67.248
Luc14 g048 1 0.056 0.03 1.849 56.099
Luc14 g048 2 0.056 0.031 1.795 56.201
Luc14 g048 3 0.06 0.034 1.782 59.902
Gal4-EED +dox g048 1 0.076 0.041 1.844 76.457
Gal4-EED +dox g048 2 0.072 0.039 1.854 71.611
Gal4-EED +dox g048 3 0.077 0.042 1.838 77.038



Diluted untreated and g046 column purified P151/158 PCR products down to 30ng/ul each.
Cut 250ng DNA with HindIII, 20 minutes at 37deg C and 20 min at 80deg C to inactivate. Did in new PCR machine.

DNA 250ng
enzyme 0.5
10xbuffer 2
water 9.17
Total 20


Dilute 1ul into 20ul water. run 1ul on bioanalyzer.