2407 pCAMBIA-Miraculin: Difference between revisions
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Latest revision as of 13:20, 1 April 2015
Cloning of 2407
[edit]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.
[edit]
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.
