MS 5 25 2017
Use IPTG to induce 483 Tth Polymerase /1464 T7 DNA Ligase / 1946 TdT bovine / 1497 ToughTaq for expression screening. Spin down cells, wash with Lysis Buffer, break down through blunted needle, and incubate with 50 ul regenerated Ni beads. Wash Ni beads, prepare to run SDS-PAGE gel.
Culture 1708HTNCre in 8L LB media. Induce with IPTG.
Culture ZXPmp / 818 / 819 / 820.
Dialysis of 1715Pfu and 1315 Mth RNA Ligase. The final concentration of 1715 Pfu is 4.10 mg/ml, totally 7ml, 28.70mg. The final concentration of 1315 Mth RNA Ligase is 6.50 mg/ml, totally 18.8ml, 122.20mg.
Small scale expression screening of 1613pET28-cold-GST-3C / 1782Tth-Ligase / 2196PBCV-1-Lig / 1498TaqDNALigase / 2295yPolyA-Pol. Run SDS-PAGE gel.
| number |
1 |
2 |
3 |
4 |
5 |
6 |
7 |
8 |
9 |
10 |
11 |
| sample |
1613 P |
1782 P |
2196 P |
1498 P |
2295 P |
MW |
1613 S |
1782 S |
2196 S |
1498 S |
2295 S |
| uL |
15 |
15 |
15 |
15 |
15 |
10 |
15 |
15 |
15 |
15 |
15 |
S: use lysis buffer S to break down cell, spin down and collect supernatant.
P: use lysis buffer P to wash sediment, sse blunted needle to break pellet, spin down and collect supernatant.
Cell culture of 1782 Tth-Ligase