1626 pET28-cold-NRF2
Cloning of 1626 pET28-cold-NRF2
[edit]Purpose: Cloning of 1626 pET28-cold-NRF2
Date: 2013-1-25---2013-1-28
Notebook:0
Page:
Clone ID:1626
Experimenters: yingying
Results
[edit]PCR
[edit]
1/27/2013
Colony Screening
[edit]Date: 2013-1-
Make miniprep and digest with
Result intepretation/Summary
[edit]Conclusion
[edit]
Cloning of 1626 pET28-cold-NRF2
[edit]Purpose: Cloning of 1626 pET28-cold-NRF2
Date: 2013-1-29--2013-1-31
Notebook:0
Page:
Clone ID:1626
Experimenters: yingying
Results
[edit]PCR
[edit]
clone into v19 pET28-cold vector using Nde1 + Xho1
Colony Screening
[edit]Date: 2013-1-31
Make miniprep and digest with Nde1 + Xho1
Result intepretation/Summary
[edit]Conclusion
[edit]
Protein Expression
[edit]1625_pCold-GST-(3c)-KEAP1#Protein_Expression
Cloning of 1626 pET28-cold-NRF2
[edit]Purpose: Cloning of 1626 pET28-cold-NRF2
Date: 2013-2-27--2013-3-1
Notebook:0
Page:
Clone ID:1626
Experimenters: yingying
Results
[edit]cloning
[edit]
F2+R
NRF2-Nde1-Nhe1-F2
catagc CATatg GCTAGC atggacttggagctgccgccgccg
NRF2-Kpn1-Xho1-R tgatcgCTCGAGGGTACCctagtttttcttaacatctggcttc
Colony Screening
[edit]Date: 2013-3-1
Make miniprep and digest with Nde1 + Xho1
Result intepretation/Summary
[edit]- <br>
Conclusion
[edit]protein expression result of NRF2 clones
[edit]| clone ID |
expression |
gel reslut |
|
| 1626, pET28-cold-NRF2 |
no |
1625 1626 1656 |
|
| 1661,pT7A3-MBP-(3c)-NRF2 |
yes, |
1661 1674 1683 |
|
| 1674,pCold-GST-(3c)-NRF2 |
need to use GST beads |
1661 1674 1683 |
|
| 1683,pCold-MBP-(3C)-NRF2 |
yes, |
1661 1674 1683 |
|
| 1698,pT7A3-NRF2 |
no |
||
| 1719,pT7A3-MBP-(E3C)-NRF2 |
no |
626 1717 1719 1720 |
|
| 1748,pCold-8His-MBP-NRF2-6His |
only MBP |
1676 1726 1708 1748 |