760 pEC2-Pme1

Cloning of760 pEC2-Pme1

Purpose: Cloning of760 pEC2-Pme1==

Date: 2014-12_2015_1

Notebook:0

Page:

Clone ID:760

Experimenters:  shanyi

Results

CLONING

gene synthesis

clone to 2015 using Nde1 + Xho1

Colony Screening

Date: 2015_1

Make miniprep, use Nde1 + Xho1 to find positive clone

Fix 760-5

pcr

fragment 1

761-PmeI-1+761-PmeI-14

fragment 2 761-PmeI-15+761-PmeI-18

clone to 2015 using Nde1 + Xho1

Colony Screening

Date: 2015_1

Make miniprep, use Nde1 + Xho1 to find positive clone

Fix 760-1

pcr

clone to 2015 using Nde1 + Xho1

Colony Screening

Date: 2015_1

Make miniprep, use Nde1 + BamH1 to find positive clone

760-fix mutation

Missing CT

Pick up 760-fix-5 as template

PCR 3-26-15

3-27-15 Repeat of 3-26

4-8-15 760 insert PCR product

760 clone

2015-4-14

760 PCR procuct digestion after innoculation