2423-pEC-pvuRts1I

Cloning of 2423-pEC-pvuRts1I

Purpose: Cloning of 2423-pEC-pvuRts1I

Name: Xin

Date: 2015 April

Results

Gene sysnthsis in two sections: 1-1 --- 1-10 ; 1-11- 1-20

The PCR about 450bp is what we interested.

  

Gel purified the inteserted PCR product, and put togeter to do the big PCR.

The PCR product is about 900bp. Purifed the PCR product from gel. and digest by Nde1/xhoI

Also digest 2015 PEC2. ligation and transformation

Screening:

Select colony#5 to sequence on 4/20/2015

Fix:

Colony#5 sequencing result have one nucleotide was wrong.

First PCR fragments: Use oligos 2423-PvuRts1I-1, and -4: 160bp

Second PCR fragements: use oligos: 2423-PvuRts1I-5, and -20: 620bp

Join two fragments together. 

Screening:

1. 2 and 3 are positive colonies.