1717-pT7A3-6H-(TEV)-MBP-(3C)

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Cloning of 1717-pT7A3-6H-(TEV)-MBP-(3C)

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Purpose: Cloning of 1717-pT7A3-6H-(TEV)-MBP-(3C)

Date: 2013-4-16---2013-4-18

Notebook:0

Page:

Clone ID:1717

Experimenters: yingying

Results

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cloning

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Colony Screening

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Date: 2013-4-18

Make miniprep and digest with Nco1 + Nde1






Result intepretation/Summary

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Conclusion

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protein screening

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lane
 1 2 3 4 5 6 7 8
 9
 10
clone ID
626,pT7A3-Taq_d289

1717,

pT7A3-6H-(TEV)-MBP-(3C)

1719,

pT7A3-MBP-(E3C)-NRF2

 1720,pT7A3-MBP-(E3C)-Keap1
 1719,

pT7A3-MBP-(E3C)-NRF2,

fraction 38
1719,

pT7A3-MBP-(E3C)-NRF2,

fraction 23

cre,

3.9ug/ul

 cre,4/2/2013
 1624,pCold-6His-GST-(E)-(3C)
host
Nico21DE3plysN
 Nico21DE3plysN
 BL21DE3pRare2pLysS
 BL21DE3pRare2pLysS
 BL21DE3pRare2pLysS
BL21DE3pRare2pLysS

Nico21DE3plysN
induction
 0.2 mM IPTG 25 oC o/n
 0.2 mM IPTG 25 oC o/n
 0.2 mM IPTG 25 oC o/n
 0.2 mM IPTG 25 oC o/n
 0.2 mM IPTG 25 oC o/n
 0.2 mM IPTG 25 oC o/n

0.2 mM IPTG 25 oC o/n
purification
 Ni beads
 Ni beads
 Ni beads
 Ni beads
 5ml HisTrap HP
 5ml HisTrap HP

Ni beads
MW
 61KD/5.97
 49KD/5.46
 112KD/4.82
 115KD/5.57
 112KD/4.82
 112KD/4.82

33KD/6.0
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