MS reports 7 10 2017-7 14 2017
07/10/2017
[edit]Culture 818 pHis-GST-His-Flag-His 8L, induced with IPTG.
Homogenize 1782 Tth-Ligase, centrifugate and load onto Ni column.
Load 1567 Exo onto QHP column.
http://excellgen.net/eln/index.php/MS_7_7_2017
Dialysis 2462 Tn5 Transposase. 1.48mg/ml, 5.5ml, purity >90%.
07/11/2017
[edit]Homogenize 818 pHis-GST-His-Flag-His, centrifugate and load onto Ni column.
Wash Ni column (load with 1782 Tth-Ligase) with Ni washing buffer/ Tris-HCl/ Buffer B. Elute 1782 Tth-Ligase with Buffer B 07/11/2017
| -0.26 |
-0.27 |
-0.28 |
-0.19 |
0.03 |
(1~5) |
| 4.68 |
6.36 |
7.35 |
5.35 |
4.74 |
(6~10) |
| 4.15 |
3.85 |
3.53 |
2.75 |
2.35 |
(11~15) |
| 1.85 |
1.53 |
1.21 |
0.54 |
0.23 |
(16~20) |
| 0.12 |
0.08 |
(21~25) |
collect 6~20.
Load 423 DeepVent-sso7d onto QHP column.
Elute 1567 T5-Exo
Run SDS-PAGE gel of 1567 T5-Exo 5.55 / 34.5 07/11/2017
|
Number |
1 |
2 |
3 |
4 |
5 |
6 |
7 |
8 |
9 |
10 |
11 |
12 |
|
sample |
19 |
20 |
21 |
22 |
23 |
24 |
25 |
MW |
26 |
27 |
28 |
30 |
|
uL |
9.1 |
5.3 |
3.9 |
3.0 |
3.1 |
2.6 |
2.4 |
10 |
3.3 |
4.8 |
5.3 |
9.1 |
|
ddH2O |
5 |
10 |
10 |
10 |
10 |
10 |
10 |
\ |
10 |
10 |
10 |
5 |
Forward lane 11~12 to the next step.
07/12/2017
[edit]Wash Ni column (load with 818 pHis-GST-His-Flag-His) with Ni washing Buffer/ Tris-HCl/ Buffer B
Elute 818 pHis-GST-His-Flag-His with Buffer B 07/12/2017
| -0.30 |
-0.26 |
0.05 |
1.95 |
2.26 |
(6~10) |
| 1.68 |
1.18 |
0.85 |
0.62 |
0.43 |
(11~15) |
| 0.30 |
0.20 |
0.14 |
0.08 |
0.05 |
(16~20) |
collect 9~16.
Load 1782 Tth-Ligase onto QHP column.
Elute 423 DeepVent-sso7d from QHP column.
Run SDS-PAGE gel of 423 DeepVent-sso7d 8.69 / 99.5 07/12/2017
|
Number |
1 |
2 |
3 |
4 |
5 |
6 |
7 |
8 |
|
sample |
14 |
15 |
16 |
17 |
18 |
MW |
20 |
22 |
|
uL |
15 |
4.6 |
2.7 |
2.7 |
4.0 |
10 |
5.0 |
5.9 |
|
ddH2O |
\ |
10 |
10 |
10 |
10 |
\ |
10 |
10 |
Forward lane 1~2 to the next step.
Dialysis of 1567 T5-Exo. 1.5mg/ml, 8.0ml.
07/13/2017
[edit]Load 818 pHis-GST-His-Flag-His onto QHP column.
Elute 1782 Tth-Ligase from QHP column.
Run SDS-PAGE gel of 1782 Tth-Ligase 6.56 / 79.1 07/13/2017
|
Number |
1 |
2 |
3 |
4 |
5 |
6 |
7 |
|
sample |
MW |
FT1 |
14 |
17 |
18 |
23 |
FT2 |
|
uL |
10 |
15 |
10 |
0.5 |
0.22 |
0.9 |
15 |
|
ddH2O |
\ |
\ |
5 |
15 |
15 |
15 |
\ |
FT1: flow through after Ni column.
FT2: flow through after QHP column.
Forward lane 5~6 to the next step.
Dialysis of 423 DeepVent-sso7d. 1.50mg/ml, 8.0ml.
07/14/2017
[edit]Dialysis of 1782 Tth-Ligase. 11.20mg/ml, 5.9ml.