MS 5 19 2017
Prepare competent cell (Nico21)
Run SDS-PAGE gel of 121-1715 Pfu and 122-1315 Mth RNA Ligase.
| 1715 Pfu 98.6 |
1315 Mth RNA Ligase 45.3 | |||||||||||
| Number |
1 |
2 |
3 |
4 |
5 |
6 |
7 |
8 |
9 |
10 |
11 |
12 |
| sample |
14 |
15 |
17 |
18 |
19 |
MW |
25 |
27 |
33 |
35 |
36 |
44 |
| uL |
0.4 |
1.0 |
0.9 |
0.4 |
0.8 |
10 |
0.9 |
1.0 |
0.5 |
0.2 |
0.2 |
0.9 |
| ddH2O |
15 |
15 |
15 |
15 |
15 |
\ |
15 |
15 |
15 | 15 |
15 |
15 |
1~5:1715
6:MW
7~12:1315
Conclusion: forward sample 14 and 15 of 1715, and 33~44 of 1315 to the next step.
Transfaction reagent 50ul/tube
QC of 1542 Native ToughTaq.
| 1 |
2 |
3 |
4 |
5 |
6 |
7 |
|
positive control |
1mg/ml |
0.1mg/ml |
10ug/ml |
1ug/ml |
0.1ug/ml |
negative control |
| 0.5ul |
0.5ul |
0.5ul |
0.5ul |
0.5ul |
0.5ul |
0 |
20ul/tube:
| dNTP |
template |
F |
R |
10x Buffer |
H2O |
Taq |
| 1ul |
0.1ul |
1ul |
1ul |
2ul |
14.4ul |
0.5ul |
Conclusion: need to do again with less cycle.