03/22/2015
Column purified unt and g032 treated nested PCR with P149/174. Will set up hybridization of 400ng PCR product in 15ul reaction:
Cell type
|
treatment
|
rep
|
260
|
280
|
260/280
|
ng/µL
|
ul for 400ng
|
ul water
|
ul hyb buff
|
total
|
Luc14 |
Unt |
1 |
0.036 |
0.02 |
1.805 |
36.09 |
11.08 |
2.42 |
1.5 |
15
|
Luc14 |
Unt |
2 |
0.042 |
0.024 |
1.801 |
42.349 |
9.45 |
4.05 |
1.5 |
15
|
Luc14 |
Unt |
3 |
0.038 |
0.021 |
1.815 |
38.219 |
10.47 |
3.03 |
1.5 |
15
|
Gal4-EED+dox |
Unt |
1 |
0.045 |
0.025 |
1.777 |
45.01 |
8.89 |
4.61 |
1.5 |
15
|
Gal4-EED+dox |
Unt |
2 |
0.042 |
0.023 |
1.836 |
42.28 |
9.46 |
4.04 |
1.5 |
15
|
Gal4-EED+dox |
Unt |
3 |
0.038 |
0.022 |
1.77 |
38.201 |
10.47 |
3.03 |
1.5 |
15
|
Luc14 |
g032 |
1 |
0.035 |
0.018 |
1.982 |
35.035 |
11.42 |
2.08 |
1.5 |
15
|
Luc14 |
g032 |
2 |
0.04 |
0.022 |
1.787 |
39.676 |
10.08 |
3.42 |
1.5 |
15
|
Luc14 |
g032 |
3 |
0.042 |
0.024 |
1.785 |
42.176 |
9.48 |
4.02 |
1.5 |
15
|
Gal4-EED+dox |
g032 |
1 |
0.043 |
0.023 |
1.851 |
42.765 |
9.35 |
4.15 |
1.5 |
15
|
Gal4-EED+dox |
g032 |
2 |
0.037 |
0.022 |
1.65 |
36.658 |
10.91 |
2.59 |
1.5 |
15
|
Gal4-EED+dox |
g032 |
3 |
0.038 |
0.022 |
1.742 |
37.775 |
10.59 |
2.91 |
1.5 |
15
|
Annealing reaction in Surveyor protocol
1ul surveyor enzyme, 1ul enhancer, 60 mins at 42deg
Dilute 1:10 in ES, run on bioanalyzer
|