MS 6 14 2017
Spin down 2295 yPolyA.
Resuspend 1483 T4 DNA Ligase, homogenize, load onto Ni column.
Wash Ni column (bind with 1498 Taq DNA Ligase) with Ni washing buffer, Tris-HCl, Buffer B, dilute and load onto QHP column.
Elute 1498 Taq DNA Ligase with Buffer B 06/14/2017
| 0.04 |
0.08 |
0.95 |
1.25 |
1.98 |
(1~5) |
| 2.84 | 4.72 |
2.16 |
1.15 |
0.78 |
(6~10) |
| 0.57 |
0.41 |
0.20 |
0.16 |
0.05 |
(11~15) |
collect 3~12.
Elute QHP column (load with 1497 ToughTaq)
Resuspend 1483 T4 DNA Ligase, homogenize, load onto Ni column.
Run SDS-PAGE gel of 1946 TdT bivine 7.67/45.7 06/14/2017
|
Number |
1 |
2 |
3 |
4 |
5 |
6 |
7 |
8 |
9 |
10 |
11 |
12 |
|
sample |
18 |
19 |
21 |
23 |
MW |
24 |
26 |
27 |
28 |
31 |
33 |
34 |
|
uL |
5.6 |
4.4 |
1.0 |
1.7 |
10 |
1.3 |
0.7 |
0.9 |
0.9 |
2.5 |
4.8 |
6.7 |
|
ddH2O |
10 |
10 |
15 |
15 |
\ |
15 |
16 |
15 |
15 |
10 |
10 |
10 |
Forward sample 18~21 to the next step.
Run SDS-PAGE gel of 1518 T3 Ligase 5.62/41.6 1601 6His-GST-i3C 6.41/50.8 1765 T4 DNA Polymerase 6.19/105.9 1942 phi29 5.72/70.6 06/14/2017
|
Number |
1 |
2 |
3 |
4 |
5 |
6 |
7 |
8 |
9 |
10 |
11 |
12 |
|
sample |
1518 S |
1601 S |
1765 S |
1942 S |
MW |
1518 P |
1601 P |
1765 P |
1942 P |
1518 U |
1601 U |
1765 U |
|
uL |
15 |
15 |
15 |
15 |
10 |
15 |
15 |
15 |
15 |
15 |
15 |
15 |
S:supernatant
P:sediment
U:uninduced
Only 1518 T3 Ligase and 1942 phi29 can do protein expression.