153 pLVX-Plus
Jump to navigation
Jump to search
Subcloning of 153 pLVX-Plus
[edit]Purpose: Subcloning of 153 pLVX-Plus
Date: August 15, 2012
Notebook:
Page:
Clone ID: 153
Experimenters: Paul Xu and Jennifer Truong
Results
[edit]Cloning
[edit]cut vector 90 pLVX-puro-HA-AKR1C4 with Xho1+BamH1, purify vector
mix equal vol of pLVXPuro-F and pLVXPuro-R, heat at 95 oC for 5 min
Slowly cool to RT, mix with vector and ligate.
pLVXPuro-F
TCGAGGGCCGCTGCGGCCTTCGAATTCTGCAGTCGACGGTAGGCCAAACAGGCCG
CCCGGCGACGCCGGAAGCTTAAGACGTCAGCTGCCATCCGGTTTGTCCGGCCTAG
pLVXPuro-R
GATCCGGCCTGTTTGGCCTACCGTCGACTGCAGAATTCGAAGGCCGCAGCGGCCC
Colony Screening
[edit]Date: August 15, 2012
Digested MiniPrep with RE Nde1 + EcoR1 to cut out CMV promoter
All samples positive for insert, used sample 1 for subcloning (all plasmids are inside Jennifer's box)
Result intepretation/Summary
[edit]
Conclusion
[edit]