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




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