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.
|