1813-pT7A3-HTNFlp-Ecoli
Jump to navigation
Jump to search
Cloning of 1813-pT7A3-HTNFlp-Ecoli
[edit]Purpose: Cloning of 1813-pT7A3-HTNFlp-Ecoli
Date: 2013-9-11---2013-9-12
Notebook:0
Page:
Clone ID:1813
Experimenters: yingying
Results
[edit]cloning
[edit]
| Lane |
1 |
2 |
3 |
| 1812,pET28b-HTNFlp-Ecoli |
1279,pT7A3 |
1512,pT7A3-Phi29DNAPol | |
| Nco1 + Xho1 | |||
Colony Screening
[edit]Date: 2013-9-12
Make miniprep and digest with Nco1 + Xho1
clone 1,2,3,4,6 right
Protein Purification
[edit]9/29/2013
Purification of Nde1 using 25 ml heparin HyperD column (XK16).
Dilute partial purified Nde1 with 50 mM Tris-HCl. Load to heparin column.
Elute with 250 ml linear gradient of 0.1 M to 0.6 M NaCl. Nde1 is eluted at 0.4 M NaCl.
| Lane |
Clone ID |
Desc |
| 1 |
1710 |
Nde1, Eluted from Heparin column |
| 2 |
1710 |
Nde1, Eluted from Heparin column |
| 3 |
1710 |
Nde1, Eluted from Heparin column |
| 4 |
1813 |
FLP, 50 mM I wash from Ni column |
| 5 |
1813 |
FLP, 300 mM I eluted from Ni, diluted |
| 6 |
1710 |
Nde1, Eluted from Heparin column |
| 7 |
1813 |
FLP, 300 mM I eluted from Ni, diluted |
| 8 |
1813 |
FLP, heparin column FT |
| 9 |
1813 |
FLP, 50 mM I wash from Ni column |
| 10 |
1835 |
MBP-Taq-Muts-6XHis, partial purified from Ni column |
| 11 |
1813 |
FLP heparin column elute |
| M |
Fermentas PageRuler Plus | |
| 12 |
1813 |
FLP heparin column elute |
| 13 |
1813 |
FLP heparin column elute |