2180 2130 2174 cre 2108s 2108u
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Expression screening, 4/13/2014, by Shanyi
? mM IPTG induction o/n at RT, 15 ml culture
g
| Lane |
Clone ID |
Name |
Competent Cells |
Sample |
Expression |
Note |
| 1 |
2180 |
pT7A3-R9-HA-iFlp |
Nico(DE3)pLysN |
S |
||
| 2 |
2180 |
Nico(DE3)pLysN |
P |
IB?9.65 /54.8 |
||
| 3 |
2180 |
pT7A3-R9-HA-iFlp |
Tuner(DE3)pLysN |
S |
||
| 4 |
2180 |
Tuner(DE3)pLysN |
P |
IB?9.65 /54.8 |
||
| M |
Fermentas |
PageRuler Plus |
||||
| 5 |
2130 |
pT7A3-6H-NLS-Cre |
Tuner(DE3)pLysN |
yes 9.87 /40.3 |
||
| 6 |
2130 |
pT7A3-6H-NLS-Cre |
Nico(DE3)pLysN |
yes 9.87 /40.3 |
||
| 7 |
2174 |
pCold-R9-HA-Cre |
Nico(DE3)pLysN |
low 10.09/44.7 |
||
| 8 |
2174 |
pCold-R9-HA-Cre |
Tuner(DE3)pLysN |
No |
||
| 9 |
Cre |
10.30 / 42.8 |
size is a little smaller | |||
| 10 |
2108 |
pCold-Tat-NLS-iFLP-6H |
s |
9.77 / 53.3 |
size is a little larger | |
| 11 |
2108 |
p |
Lyse cell by freeze-thawing, 2 cycles, using cell lysis buffer, spin--> Supernatant (S)
Add screening lysis buffer containing 4M urea, 1% SDS in Buffer A to pellet, solubilize and spin-->P