MS reports 7 3 2017-7 7 2017
07/03/2017
[edit]Culture 1567 Exo 200ml.
Culture Tn5 transposase 8L, induced with IPTG.
Homogenize 1811 vPol, load onto Ni column.
Load 1458 KOD onto QHP column.
Ship order 4890 and 4891.
Prepare SDS-PAGE Gel staining.
07/04/2017
[edit]Culture 423 pET30b-DeepVent-sso7d 200ml.
Culture 1567 Exo 8L, induced with IPTG.
Homogenize 2462 Tn5 transposase, load onto Ni column.
Load 1995 Kfu onto QHP column.
Wash Ni column (load with 1811 vPol) with Ni washing buffer/ Tris-HCl/ Buffer B.
Elute 1811 vPol with Buffer B 07/04/2017
| -0.02 |
-0.11 |
-0.14 |
-0.21 |
-0.08 |
(1~5) |
| 1.18 |
3.41 |
6.35 |
17.97 |
36.35 |
(6~10) |
| 20.81 |
15.44 |
10.71 |
4.83 |
1.61 |
(11~15) |
| 0.76 |
0.32 |
0.24 |
0.12 |
0.08 |
(16~20) |
collect 6~17.
Elute 1458 KOD
Run SDS-PAGE gel of 1458 KOD 7.91/92.2 07/04/2017
|
Number |
1 |
2 |
3 |
4 |
5 |
6 |
7 |
8 |
9 |
10 |
11 |
12 |
|
sample |
16 |
18 |
20 |
22 |
24 |
26 |
28 |
30 |
MW |
32 |
34 |
36 |
|
uL |
10 |
0.2 |
0.1 |
0.13 |
0.13 |
0.2 |
0.23 |
0.7 |
10 |
2.0 |
4.2 |
7.7 |
|
ddH2O |
5 |
15 |
15 |
15 |
15 |
15 |
15 |
15 |
\ |
15 |
10 |
5 |
forward lane 1~3 to the next step.
07/05/2017
[edit]Culture 1782 Tth Ligase 200ml.
Culture 423 pET30b-DeepVent-sso7d 8L, induced with IPTG.
Homogenize 1567 Exo, load onto Ni column.
Load 1811 vPol onto QHP column.
Wash Ni column (load with 2462 Tn5 transposase) with Ni washing buffer/ Tris-HCl/ Buffer B.
Elute 2462 Tn5 transposase with Buffer B 07/05/2017
| -0.24 |
-0.26 |
-0.24 |
-0.23 |
-0.20 |
(6~10) |
| -0.15 |
0.28 |
1.31 |
0.38 |
0.04 |
(11~15) |
| 0.00 |
(16~20) |
collect 12~15.
Elute 1995 Kfu
Run SDS-PAGE gel of 1995 Kfu 7.87 / 99.3 07/05/2017
|
Number |
1 |
2 |
3 |
4 |
5 |
6 |
7 |
8 |
9 |
10 |
11 |
12 |
|
sample |
17 |
18 |
19 |
20 |
21 |
22 |
23 |
MW |
25 |
27 |
29 |
31 |
|
uL |
9.1 |
1.5 |
0.8 |
0.7 |
1.1 |
1.7 |
2.4 |
10 |
3.3 |
4.4 |
6.7 |
11.1 |
|
ddH2O |
5 |
15 |
15 |
15 |
15 |
15 |
15 |
\ |
10 |
10 |
10 |
5 |
forward lane 1~4 to the next step.
Dialysis of 1458 KOD. 14.27mg/ml, 4.5ml, purity > 90%
07/06/2017
[edit]Culture 1782 Tth Ligase 8L, induced with IPTG.
Homogenize 423 pET30b-DeepVent-sso7d, load onto Ni column.
Load 2462 Tn5 transposase onto QHP column.
Wash Ni column (load with 1567 Exo) with Ni washing buffer/ Tris-HCl/ Buffer B.
Elute 1567 Exo with Buffer B 07/06/2017
| -0.25 |
-0.26 |
-0.25 |
-0.25 |
-0.23 |
(1~5) |
| -0.10 |
1.82 |
1.04 |
0.36 |
0.15 |
(6~10) |
| 0.07 |
0.04 |
0.03 |
0.01 |
0.01 |
(11~15) |
collect 7~11.
Elute 1811 vPol
Run SDS-PAGE gel of 1811 vPol 6.63 / 69.2 07/06/2017
|
Number |
1 |
2 |
3 |
4 |
5 |
6 |
7 |
8 |
9 |
10 |
11 |
12 |
|
sample |
8 |
10 |
12 |
14 |
15 |
16 |
17 |
MW |
18 |
19 |
20 |
21 |
|
uL |
5.9 |
3.9 |
4.0 |
2.1 |
0.23 |
0.38 |
0.3 |
10 |
0.44 |
0.55 |
0.7 |
0.1 |
|
ddH2O |
10 |
10 |
10 |
15 |
15 |
15 |
15 |
\ |
15 |
15 |
15 |
15 |
forward lane 1~7 to the next step.
Dialysis of 1995 Kfu. 4.18mg/ml, 4.5ml, purity > 90%
07/07/2017
[edit]Wash Ni column (load with 423 DeepVent-sso7d) with Ni washing buffer/ Tris-HCl/ Buffer B.
Elute 423 DeepVent-sso7d with Buffer B 07/07/2017
| -0.29 |
-0.26 |
-0.14 |
0.99 |
1.27 |
(6~10) |
| 0.94 |
0.80 |
0.70 |
0.64 |
0.61 |
(11~15) |
| 0.57 |
0.50 |
0.49 |
0.44 |
0.40 |
(16~20) |
| 0.39 |
0.37 |
0.30 |
0.28 |
0.24 |
(21~25) |
| 0.22 |
0.22 |
0.16 |
0.11 |
0.08 |
(26~30) |
collect 9~27.
Elute 2462 Tn5 transposase
Run SDS-PAGE gel of 2462 Tn5 transposase 8.41 / 83.8 07/07/2017
|
Number |
1 |
2 |
3 |
4 |
5 |
6 |
7 |
8 |
9 |
10 |
|
sample |
22 |
23 |
24 |
25 |
26 |
MW |
27 |
28 |
29 |
30 |
|
uL |
6.3 |
8.4 |
7.2 |
5.6 |
5.3 |
10 |
6.3 |
7.2 |
8.4 |
10 |
|
ddH2O |
10 |
5 |
5 |
10 |
10 |
\ |
10 |
5 |
5 |
5 |
forward lane 1~4 to the next step.
Dialysis of 1811 vPol. 36.0mg/ml, 10.0ml, purity > 90%