1542-pT7A3-sso7d-Native-ToughTaq
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Cloning of 1542-pT7A3-sso7d-Native-ToughTaq
[edit]Purpose: Cloning of 1542-pT7A3-sso7d-Native-ToughTaq
Date: 2013-3-5---2013-3-7
Notebook:0
Page:
Clone ID:1542
Experimenters: yingying
Results
[edit]subcloning
[edit]
Colony Screening
[edit]Date: 2013-3-7
Make miniprep and digest with Nde1 + Sal1
Result intepretation/Summary
[edit]Conclusion
[edit]protein screen
[edit]
| lane |
name |
clone ID |
host |
induction |
purification |
MW |
| 1 |
pT7A3-sso7d-Native-ToughTaq |
1542 |
4 ml,
BL21(DE3)Star, pRare2-pLysS |
0.2 mM IPTG 25 oC o/n |
Ni beads |
62KD |
| 2 |
pT7A3-VVCap-L |
1544 |
4 ml,
BL21(DE3)Star, pRare2-pLysS |
0.2 mM IPTG 25 oC o/n |
Ni beads |
34KD |
| 3 |
pT7A3-VVCap |
1545 |
4 ml,
BL21(DE3)Star, pRare2-pLysS |
0.2 mM IPTG 25 oC o/n |
Ni beads |
34KD |
| 4 |
pT7A3-SOD |
1627 |
4 ml,
BL21(DE3)Star, pRare2-pLysS |
0.2 mM IPTG 25 oC o/n |
Ni beads |
16KD |
| 5 |
pCold-MBP-SM2 |
1679 |
4 ml,NiCO21(DE3)pLysN, |
0.2 mM IPTG 25 oC o/n |
Ni beads |
74KD |
| 6 |
pCold-GST-wtSM2 |
1680 |
4 ml,NiCO21(DE3)pLysN, |
0.2 mM IPTG 25 oC o/n |
get lysis supernatant to run gel |
94KD |
QC
[edit]2013-8-15
15ul elution in 300 mM Imidozale, 20 mM Tris-HCl pH 8.0, 600 mM NaCl
PCR test