MS reports 6 19 2017-6 23 2017
06/19/2017
[edit]Plasmid transformation of 1613 pET28-cold-GST-3C and 1697 pCold-GST-(3c)-wt3C.
load 2295 yPolyA onto Ni column.
06/20/2017
[edit]wash 1483 T4 DNA Ligase and 2295 yPolyA from Ni column.
Culture 1613 pET28-cold-GST-3C 200 ml.
Spin down 1518 T3 ligase and homogenize, load onto Ni column.
06/21/2017
[edit]Culture 1697 pCold-GST-(3c)-wt3C 200ml.
Culture 1613 pET28-cold-GST-3C 8L, induced with IPTG at 16oC.
Homogenize 1942 phi29, load onto Ni column.
Elute QHP column (load with 1483 T4 DNA Ligase)
Run SDS-PAGE gel of 1483 T4 DNA Ligase 6.63 / 57.3 06/21/2017
|
Number |
1 |
2 |
3 |
4 |
5 |
6 |
7 |
8 |
|
sample |
10 |
11 |
12 |
13 |
14 |
15 |
16 |
MW |
|
uL |
8.4 |
0.16 |
0.13 |
0.6 |
1.2 |
2.5 |
7.2 |
10 |
|
ddH2O |
5 |
15 |
15 |
15 |
15 |
10 |
5 |
\ |
forward lane 1~3 to the next step.
Dilute 2295 yPolyA and load onto QHP column.
Elute QHP column (load with 2295 yPolyA)
Miniprep. 2462 Tn5 transposase.
Plasmid transformation of 818 N-6His, Internal 6His. Flag, C-6His / 819 N-His, GST, Myc, C-His / 820 N-6His, GST, HA, C-6XHis / 1708 HTNCre / 2462 Tn5 transposase
06/22/2017
[edit]Culture 1518 T3 ligase 200ml.
Culture 1697 pCold-GST-(3c)-wt3C 8L, induced with IPTG at 16oC.
Spin down and homogenize 1613 pET28-cold-GST-3C 8L, load onto Ni column.
Wash Ni column (load with 1942 phi29) with Ni washing Buffer, Tris-HCl, Buffer B.
Elute QHP column (load with 1518 T3 ligase)
Run SDS-PAGE gel of 1518 T3 ligase 5.62 / 41.6 06/22/2017
|
Number |
1 |
2 |
3 |
4 |
5 |
6 |
7 |
8 |
9 |
10 |
|
sample |
11 |
12 |
13 |
14 |
15 |
16 |
17 |
18 |
19 |
MW |
|
uL |
15 |
15 |
15 |
15 |
15 |
15 |
15 |
15 |
15 |
10 |
forward lane 7~8 to the next step.
Run SDS-PAGE gel of 2295 yPolyA 7.63 / 66.2 06/22/2017
|
Number |
1 |
2 |
3 |
4 |
5 |
6 |
7 |
8 |
|
sample |
20 |
21 |
22 |
23 |
24 |
25 |
26 |
MW |
|
uL |
4.6 |
0.8 |
0.5 |
1.0 |
1.2 |
1.3 |
1.5 |
10 |
|
ddH2O |
10 |
15 |
15 |
15 |
15 |
15 |
15 |
\ |
forward lane 1~3 to the next step.
06/23/2017
[edit]Spin down 1697 pCold-GST-(3c)-wt3C.
Dialysis.
1483 T4 DNA Ligase, 32mg/ml, 2.0ml, purity >90%.
2295 yPolyA, 4.7mg/ml, 2.0ml, purity >90%.
1518 T3 ligase, 0.36mg/ml, 12ml, purity 90%.