MS reports 5 30 2017-6 2 2017
05/30/2017
[edit]Culture 1782 Tth Ligase (8L), induced with IPTG, over night.
Culture 483 Tth Polymerase (200ml), over night.
Prepare Ni Elution Buffer (50mM phosphate buffer, pH7.0, 250mM NaCl, 100mM Arginine, 300mM imidazole, 10% glycerol)
Wash Ni column (bind with 1708 HTN Cre) with Ni washing Buffer, phosphate buffer and Ni Elution buffer.
Prepare SDS-PAGE Gel Stain.
Prepare Buffer A2, SP Dilution Buffer, Ni Elution Buffer.
05/31/2017
[edit]Culture 483 Tth Polymerase (8L), induced with IPTG.
Spin down 1782 Tth ligase, homogenize, centrifugation, load onto Ni column.
Prepare LB media.
Prepare SP Washing Buffer and SP Elution Buffer.
Elute SP FF column (bind with 1708 HTN Cre) with SP washing buffer and SP Elution buffer. Dialysis over night.
Culture 1708 HTN Cre (200ml).
06/01/2017
[edit]Culture 1708 HTN Cre (8L), induced with IPTG, over night.
Spin down 483 Tth Polymerase.
Dialysis of 1708 HTN Cre, total 37ml, 1.07mg/ml. Need to further dialyse.
Culture 1464 T7 DNA ligase (200ml)
Wash Ni column (bind with 1782 Tth ligase) with Ni washing Buffer/ Tris-HCl/ Buffer B,
Elute 1782 Tth ligase with Buffer B 06/01/2017
| -0.08 |
0.12 |
1.07 |
3.23 |
5.68 |
(1~5) |
| 3.70 |
2.49 |
1.46 |
0.87 |
0.59 |
(6~10) |
| 0.43 |
0.31 |
0.30 |
0.19 |
0.11 |
(11~15) |
load sample 3~12 onto QHP column.
06/02/2017
[edit]Regenerate Ni column and SP FF column.
Elute 1782 Tth Ligase.
Homogenize 483 Tth Polymerase, spin down, load supernatant onto Ni column.
Deal with order 4874, ship to Neitherlands.