1727 pT7A30

Cloning of 1727 pT7A30

Purpose: Cloning of 1727 pT7A30

Date: 2013-5-6

Notebook:0

Page:

Clone ID:1727

Experimenters: shanyi

Results

cloning

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

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.