1727 pT7A30
Cloning of 1727 pT7A30
[edit]Purpose: Cloning of 1727 pT7A30
Date: 2013-5-6
Notebook:0
Page:
Clone ID:1727
Experimenters: shanyi
Results
[edit]cloning
[edit]digest pT7A3 vector or any pT3A3 clone with Xba1 + Xho1---> cut vector out
digest pET30b with Xba1 + Xho1--> cut out 232 bp MCS fragment
ligate and transform
Colony Screening
[edit]Date: 2013-3-
Make miniprep and digest with Kpn1 + Sma1, should get one band (the parenta vector pET28b does not have Kpn1 site, and there is one Kpn1 site in pET30b)
1727digest_with_XbaIand_XhoI
Miniprep of 945pMiniT2_947pMiniT2_3131_1727_1834
945pMiniT2:1-4 EcoRI
947pMiniT2:5-8 n EcoRI
3131:9-13 BssHII+XhoI
1727:14 XbaI+NcoI
1834:15 XbaI+NcoI
Buffer:Fastdigest
Time:30Min
Results:945pMiniT2 and 947pMiniT2 are good, so sent them to do sequencing. 3131 is not good, 1727 has no band, 1834 has one band.
Result intepretation/Summary
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Conclusion
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