09/08/2015
Purified PCR products, set up annealing reactions for 24h time point
Cell type
|
drug
|
gRNA
|
time post xfect
|
rep
|
260
|
280
|
260/280
|
ng/µL
|
ul for 400ng
|
ul ann buff
|
ul water
|
Gal4 |
dox |
g034 |
24 |
1 |
0.076 |
0.042 |
1.813 |
76.239 |
5.25 |
1.50 |
8.25
|
Gal4 |
dox |
g034 |
24 |
2 |
0.087 |
0.048 |
1.812 |
87.35 |
4.58 |
1.50 |
8.92
|
Gal4 |
dox |
g034 |
24 |
3 |
0.08 |
0.043 |
1.851 |
80 |
5.00 |
1.50 |
8.50
|
Gal4 |
puro |
g034 |
24 |
1 |
0.085 |
0.048 |
1.788 |
85.425 |
4.68 |
1.50 |
8.82
|
Gal4 |
puro |
g034 |
24 |
2 |
0.05 |
0.029 |
1.725 |
50.174 |
7.97 |
1.50 |
5.53
|
Gal4 |
puro |
g034 |
24 |
3 |
0.078 |
0.044 |
1.783 |
77.729 |
5.15 |
1.50 |
8.35
|
Ran nested PCR for the remaining samples for the time course experiment. 1:1000 dilution, P149/176, staggered the set up annealing reactions and surveyor for 48h and 72h, ran on BA:
Cell type
|
drug
|
gRNA
|
time post xfect
|
rep
|
260
|
280
|
260/280
|
ng/µL
|
ul for 400ng
|
ul ann buff
|
ul water
|
Gal4 |
dox |
g034 |
48 |
1 |
0.087 |
0.049 |
1.789 |
87.049 |
4.60 |
1.50 |
8.90
|
Gal4 |
dox |
g034 |
48 |
2 |
0.089 |
0.049 |
1.808 |
88.656 |
4.51 |
1.50 |
8.99
|
Gal4 |
dox |
g034 |
48 |
3 |
0.08 |
0.043 |
1.853 |
79.666 |
5.02 |
1.50 |
8.48
|
Gal4 |
puro |
g034 |
48 |
1 |
0.08 |
0.04 |
1.972 |
79.795 |
5.01 |
1.50 |
8.49
|
Gal4 |
puro |
g034 |
48 |
2 |
0.086 |
0.045 |
1.911 |
85.877 |
4.66 |
1.50 |
8.84
|
Gal4 |
puro |
g034 |
48 |
3 |
0.069 |
0.034 |
2.024 |
68.808 |
5.81 |
1.50 |
7.69
|
Cell type
|
drug
|
gRNA
|
time post xfect
|
rep
|
260
|
280
|
260/280
|
ng/µL
|
ul for 400ng
|
ul ann buff
|
ul water
|
Gal4 |
puro |
g034 |
72 |
1 |
0.084 |
0.044 |
1.91 |
84.274 |
4.75 |
1.50 |
8.75
|
Gal4 |
puro |
g034 |
72 |
2 |
0.069 |
0.036 |
1.92 |
69.015 |
5.80 |
1.50 |
7.70
|
Gal4 |
puro |
g034 |
72 |
3 |
0.084 |
0.044 |
1.921 |
83.748 |
4.78 |
1.50 |
8.72
|
Gal4 |
dox |
g034 |
72 |
1 |
0.063 |
0.032 |
1.937 |
62.92 |
6.36 |
1.50 |
7.14
|
Gal4 |
dox |
g034 |
72 |
2 |
0.059 |
0.031 |
1.919 |
59.311 |
6.74 |
1.50 |
6.76
|
Gal4 |
dox |
g034 |
72 |
3 |
0.078 |
0.041 |
1.889 |
78.341 |
5.11 |
1.50 |
8.39
|
|