2407 pCAMBIA-Miraculin

Cloning of 2407

Purpose: Cloning of 2407 pCAMBIA-35s promoter-Miraculin-terminator

Date:  2015-3-24

Notebook: 

Page: 

Clone ID: 2412

Experimenters: Xin

subclone 2422 promoter-miraculin-terminator into vector 1153.

 

ligation, transformation

PCR screening: as the PCR cycle is 25, so no strong band can be found on on the gel photo. 

But there is a band on the gel under dark for all the colony on 400 bp location. Select the colony 2,4,and 5 mini plasmid prepation.

After analysis, found out the selection of insert was wrong. It should be1600bp instead of 700bp.

Paul redo it again.

Select 8 colonies for screening.

 

 

Reselect some colonies to screen, by PCR the primers are:

2405-Miraculin-Term-Kpn1-F	

pCAMBIA-2301-Nos-seq-R

The colony # 4, 5, 6,10,11,12 16 have the right size product -600bp.