1673-pET28b-NdeI
Cloning of 1673-pET28b-NdeI
[edit]Purpose: Cloning of 1673-pET28b-NdeI
Date: 2013-3-4---2013-3-7
Notebook:0
Page:
Clone ID:1673
Experimenters: yingying
Results
[edit]cloning
[edit]
pcr1+pcr2
Colony Screening
[edit]Date: 2013-3-7
Make miniprep and digest with Nde1+Xho1
Result intepretation/Summary
[edit]- <br>
Conclusion
[edit]<br>
<br>
<br>
<br>
Cloning of 1673-pET28b-NdeI
[edit]Purpose: Cloning of 1673-pET28b-NdeI
Date: 2013-3-11---2013-3-14
Notebook:0
Page:
Clone ID:1673
Experimenters: yingying
Results
[edit]cloning
[edit]
fix the problem:
PCR using
Fragment 1
i_NdeI1-1+i_NdeI2-10
Fragment 2
i_NdeI2-11 + i_NdeI2-14
Gel purify, then join two fragments using
i_NdeI1-1 +i_NdeI2-14
Colony Screening
[edit]Date: 2013-3-14
Make miniprep and digest with Nde1+Xho1
Result intepretation/Summary
[edit]Conclusion
[edit]protein screening
[edit]
| lane |
name |
clone ID |
host |
induction |
purification |
MW | |
| 1 |
pT7A3-T5-Exo |
1567 |
4 ml,NiCO21(DE3)pLysN |
0.2 mM IPTG 25 oC o/n |
Ni beads |
21KD/5.95 | |
| 2 |
pCold-TF-(3c) |
1614 |
4 ml,NiCO21(DE3)pLysN |
0.2 mM IPTG 25 oC o/n |
Ni beads |
55.9KD/5.19 | |
| 3 |
pET28b-Nde1 |
1673 |
4 ml,NiCO21(DE3)pLysN |
0.2 mM IPTG 25 oC o/n |
Ni beads |
44KD/8.41 | |
| 4 |
pTCold-MBP-(3C) |
1677 |
4 ml,NiCO21(DE3)pLysN |
0.2 mM IPTG 25 oC o/n |
Ni beads |
44KD | |
| 5 |
pCold-C(3C)-MBP-6H |
1692 |
4 ml,NiCO21(DE3)pLysN |
0.2 mM IPTG 25 oC o/n |
Ni beads |
46KD/5.77 | |
| 6 |
pT7A3-wtSM2 |
1694 |
4 ml,NiCO21(DE3)pLysN |
0.2 mM IPTG 25 oC o/n |
Ni beads, should use CBD beads |
33KD/6.30 | |
| 7 |
pCold-MBP-(3C)-TEV |
1696 |
4 ml,NiCO21(DE3)pLysN |
0.2 mM IPTG 25 oC o/n |
Ni beads |
72KD/6.04 | |
| 8 |
pCold-GST-(3c)-wt3C |
1697 |
4 ml,NiCO21(DE3)pLysN |
0.2 mM IPTG 25 oC o/n |
Ni beads |
21KD/8.47 | |
| 9 |
pT7A3-MBP-(E3C)-p53wt |
1707 |
4 ml,
BL21(DE3)Star, pRare2-pLysS |
0.2 mM IPTG 25 oC o/n |
Ni beads |
88KD/5.76 | |
| 10 |
pET28b-hINFA2 |
1711 |
4 ml,
BL21(DE3)Star, pRare2-pLysS |
0.2 mM IPTG 25 oC o/n |
Ni beads |
21KD/5.95 | |