2423-pEC-pvuRts1I
Cloning of 2423-pEC-pvuRts1I
[edit]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:
[edit]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:
[edit]
- 2 and 3 are positive colonies.