2410 pFastBac-VSVG-CMV5-Flp
Cloning of2410 pFastBac-VSVG-CMV5-Flp
[edit]Purpose: Cloning of2410 pFastBac-VSVG-CMV5-Flp==
Date: 2015_1
Notebook:0
Page:
Clone ID:2410
Experimenters: shanyi
Results
[edit]CLONING
[edit]PCR CMV promoter from a clone such as 2216 pCMV5-BirA
2410-CMV-Sal1-F tagaca GTCGAC GCCCATATATGGAGTTCCGCGTTAC
2410-CMV-PH-R GTTATCATTTTAATTATCTCCATAAACTTGGACCTGGGAGTGGACAC
PCR PH promoter from pFastbac vector
2410-CMV-PH-F GTGTCCACTCCCAGGTCCAAGTTTATGGAGATAATTAAAATGATAAC
2410-PH-BamH1-R (CATACCGTCCCACCATCGGGCGCGGATCCCGGTC) GACCGGGATCCGCGCCCGATGGTGGGACGGTATG
Join CMV5 and PH promoter by
2410-CMV-Sal1-F tagaca GTCGAC GCCCATATATGGAGTTCCGCGTTAC
2410-PH-BamH1-R (CATACCGTCCCACCATCGGGCGCGGATCCCGGTC) GACCGGGATCCGCGCCCGATGGTGGGACGGTATG
digest vector with Xho1 + BamH1 digest PCR product with Sal1 + BamH1
Colony Screening
[edit]Date: 2015_1
Make miniprep, use NCO1 to find positive clone,will get three bands.
