09/10/2015
Ran g031 P197/198 PCR samples on gel. No band in water-only control PCR. Gal4-EED +dox g031 rep 2 failed. Re-ran with 5ul of gDNA and it worked.
Nested PCR of g031 with P149/P174. No band in water-only control PCR.
gDNA extraction of g025 and g044 treated cells
Cells
|
drug
|
gRNA
|
rep
|
260
|
280
|
260/280
|
ng/µL
|
ul for 200
|
ul water 15-
|
Gal4-EED |
puro |
g025 |
1 |
0.015 |
0.008 |
1.973 |
15.45 |
12.94 |
2.06
|
Gal4-EED |
puro |
g025 |
2 |
0.029 |
0.015 |
1.993 |
28.969 |
6.90 |
8.10
|
Gal4-EED |
puro |
g025 |
3 |
0.02 |
0.01 |
2.032 |
20.101 |
9.95 |
5.05
|
Gal4-EED |
dox |
g025 |
1 |
0.037 |
0.019 |
2.006 |
37.202 |
5.38 |
9.62
|
Gal4-EED |
dox |
g025 |
2 |
0.08 |
0.041 |
1.974 |
80.301 |
2.49 |
12.51
|
Gal4-EED |
dox |
g025 |
3 |
0.075 |
0.039 |
1.926 |
74.544 |
2.68 |
12.32
|
Gal4-EED |
puro |
g044 |
1 |
0.015 |
0.01 |
1.522 |
14.529 |
13.77 |
1.23
|
Gal4-EED |
puro |
g044 |
2 |
0.023 |
0.012 |
1.893 |
23.315 |
8.58 |
6.42
|
Gal4-EED |
puro |
g044 |
3 |
0.05 |
0.025 |
2.008 |
49.979 |
4.00 |
11.00
|
Gal4-EED |
dox |
g044 |
1 |
0.034 |
0.016 |
2.102 |
33.619 |
5.95 |
9.05
|
Gal4-EED |
dox |
g044 |
2 |
0.069 |
0.037 |
1.881 |
69.057 |
2.90 |
12.10
|
Gal4-EED |
dox |
g044 |
3 |
0.044 |
0.023 |
1.917 |
44.009 |
4.54 |
10.46
|
Set up phusion PCR with P197/198 with 200ng gDNA
|