1150 pHis.MBP.TEV

Cloning of pHis.MBP.TEV(1150)
Purpose: to clone pHis.MBP.TEV
Date: 2012-5-1 to 2012-5-
Notebook: Xiaojing Wang
Page: 004237

Clone ID: 1150
Experimenters: Xiaojing Wang

Results

Digest pHis.MBP 1116 with Nde1 + EcoR1 --> insert
Digest pHis-GST-TEV 1030 with Nde1 + EcoR1 --> vector (discard 660bp GST fragment )

Ligate and transform TOP10
Select 4 clones to mini-prep
1150 plasmids are digested by Nde1 + EcoR1

No plasmid is correct.

repeat 5/4/2012

Ligate and transform TOP10
Select 6 clones to mini-prep
1150 plasmids are digested by Nde1 + EcoR1

No plasmid is correct.

New method 5/9/2012
digest pHis.MBP 1116 with EcoR1 + Xho1 -->vector
Digest pHis-GST-TEV 1030 with EcoR1+ Xho1--> insert

--Paul.xu 15:15, 12 May 2012 (EST)

5/12/2012.

pick 5 colonies, make miniprep, digest with Nde1 or EcoR1 + Xho1. DNA marker is Fermentas 1kb plus (20000, 10000, 7000, 5000, 4000, 3000, 2000, 1500, 1000, 700, 500, 400, 300, 200, 75).

--Paul.xu 15:15, 12 May 2012 (EST)