MS 6 29 2017
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Culture 2462 Tn5 transposase 200ml.
Culture 1811 vPol 8L, induced with IPTG.
Homogenize 1995 Kfu, load onto Ni column.
Wash Ni column (load with 1458 KOD) with Ni washing Buffer/ Tris-HCl/ Buffer B, store at -80oC.
Elute 1458 KOD with Buffer B 06/29/2017
| -0.02 |
-0.00 |
-0.01 |
0.09 |
0.12 |
(1~5) |
| 4.98 |
18.72 |
18.77 |
11.26 |
6.98 |
(6~10) |
| 4.54 |
2.84 |
1.89 |
1.31 |
0.92 |
(11~15) |
| 0.72 |
0.58 |
0.51 |
0.44 |
0.39 |
(16~20) |
| 0.24 |
0.16 |
0.13 |
0.07 |
0.04 |
(21~25) |
collect 6~21.
Elute 1518 T3 ligase from QHP column.
Run SDS-PAGE gel of 1518 T3 ligase 5.62/41.6 06/29/2017
|
Number |
1 |
2 |
3 |
4 |
5 |
6 |
7 |
8 |
9 |
10 |
11 |
12 |
|
sample |
26 |
28 |
29 |
30 |
31 |
32 |
33 |
34 |
MW |
35 |
36 |
37 |
|
uL |
6.7 |
4.8 |
4.2 |
3.4 |
3.5 |
2.5 |
2.0 |
1.8 |
10 |
1.9 |
2.6 |
3.7 |
|
ddH2O |
10 |
10 |
10 |
10 |
10 |
10 |
15 |
15 |
\ |
15 |
10 |
10 |
forward lane 10~12 to the next step.