1454 pEC

Cloning of 1454 pEC

Purpose: pEC

Date: 2012-11-8

Notebook: 

Page:

Clone ID: 1454

Experimenters: Shanyi

Results

11/7/2012, Digestion

Clone 1.2 is pCR2.1 EcoR1 digest and religate.

1.6 clone 1 and 3 are not good, they are two copies of Amp-(or Amp-Kan) ligated from Alwn1-DraIII fragment from pCR2.1

2.1 clone #4 is pRSetA.

2.1 Clone # 5 and 6 are from pRSET-A

11-05-2012

subcloning

Digest 1457 pCR2.1 with DraIII + AlwN1---vector, cut out 1296 bp fragment
digest pET28b-amp with DraIII + AlwN1---insert,
ligate and transform, plate on LB/amp

Colony Screening

digest with \DraIII + AlwN1

first 1454(from first step) digest with HindIII + EcoR1

the second 1454 (from first step) digest with Pst1

Colony Screening

Date: 2012-

Make miniprep and digest with DraIII + Nco1.