1755 pCold-6H
Jump to navigation
Jump to search
Cloning of 1755 pCold-6H
[edit]Purpose: Cloning of1755 pCold-6H
Date: 2013-6--7-
Notebook:0
Page:
Clone ID:1755
Experimenters: Shanyi
Results
[edit]cloning
[edit]PCR cold promoter from 1729 pCold-TF-(E3C)-3C using
Cold-BglII-F GGATCGAGATCTGCCGATTAATCATAAATATG
Cold-EcoR1-R
(atagCACC ATG ggc AAT CAC AAA GTG CAT CAT CAT CAT CAT CAT ATg ggtacc ggatcc gaattc GGTCGa)
tCGACCgaattcggatccggtacccATATGATGATGATGATGATGCACTTTGTGATTgccCATGGTGctat
72oC 25s, 24cycle, Q5 HOT START POLYMERASE
Digest vector v19 pET28-cold with BglII + EcoR1
Digest purified PCR product with BglII + EcoR1
ligate and transform NEB turbo
Colony Screening
[edit]Date: 2013-6-7
Make miniprep and digest with BglII + EcoR1
