MS 6 28 2017
Culture 1811 VPol 200ml.
Culture 1995 Kfu 8L, induced with IPTG.
Homogenize 1458 KOD, load onto Ni column.
Elute 1518 T3 ligase with Buffer B 06/28/2017
| -0.22 |
-0.20 |
-0.18 |
0.09 |
1.22 |
(1~5) |
| 0.97 |
0.75 |
0.55 |
0.41 |
0.31 |
(6~10) |
| 0.22 |
0.15 |
0.11 |
0.05 |
0.03 |
(11~15) |
collect 5~11.
Load 1518 T3 ligase onto QHP column.
Elute 1697 GST-wt3C
Run SDS-PAGE gel of 1942 phi 29 5.72/70.6 06/28/2017
|
Number |
1 |
2 |
3 |
4 |
5 |
6 |
|
sample |
11 |
12 |
13 |
14 |
15 |
MW |
|
uL |
12.5 |
4.0 |
4.2 |
5.3 |
8.4 |
10 |
|
ddH2O |
\ |
10 |
10 |
10 |
5 |
\ |
forward lane 1~2 to the next step.
Small scale screening of 2462 Tn5 transposase 8.41 / 83.8 818 N-6His 6.12 / 31.9 819 N-His 5.92 / 31.2 820 N-6His 5.99 / 31.2
Run SDS-PAGE gel 06/28/2017
|
Number |
1 |
2 |
3 |
4 |
5 |
|
sample |
MW |
2462 |
818 |
819 |
820 |
|
uL |
10 |
15 |
15 |
15 |
15 |
2462 818 819 820 can do protein purification.
Run SDS-PAGE gel of 1697 GST-wt3C 8.48/21.4 06/28/2017
|
Number |
1 |
2 |
3 |
4 |
5 |
6 |
7 |
8 |
9 |
10 |
11 |
12 |
|
sample |
18 |
19 |
20 |
22 |
23 |
MW |
24 |
25 |
26 |
27 |
29 |
31 |
|
uL |
1.3 |
0.9 |
0.9 |
0.8 |
0.8 |
10 |
1.0 |
1.4 |
1.8 |
2.4 |
4.6 |
7.7 |
|
ddH2O |
15 |
15 |
15 |
15 |
15 |
\ |
15 |
15 |
15 |
10 |
10 |
5 |
forward lane 1~2 to the next step.