2228 pFastBac-Dual-VSVG-HIVGag-Cre-CD63-Cre
Cloning of 2228 pFastBac-Dual-VSVG-HIVGag-Cre-CD63-Cre
[edit]Purpose: Cloning of 2228 pFastBac-Dual-VSVG-HIVGag-Cre-CD63-Cre
Date: 2014-6
Notebook:0
Page:
Clone ID:2228
Experimenters: shanyi
Results
[edit]cloning
[edit]1. PCR CD63-Cre from 2119 pCMV5-CD63-iCre using
p10-Sal1-CD63-F
ATTTACAATCACTCGACGAAGACTGTCGACATGGCGGTGGAAGGAGGAATG
iCre-Xba1-R tagctaTCTAGACTAATCGCCATCTTCCAGCAG
2. PCR p10 promoter from pFastbac-Dual
p10-Not1-F gctac GCGGCCGC GAGTATACGGACCTTTAATTC
p10-Sal1-CD63-R
(CATTTTATTTACAATCACTCGACGAAGACTGTCGAC)
GTCGACAGTCTTCGTCGAG
TGATTGTAAATAAAATG
3. Join gel 2 fragments (maybe it is NOT necessary to purify two fragment on gel) using
p10-Not1-F + Flpe-Xba1-R
4. clone into Not1 + Xba1 digested 2226 pFastBac-Dual-VSVG-HIVGag-FLP
Colony Screening
[edit]Date: 2014-6
Colony Screening
[edit]Date: 2014-6
mini prep and digest with EcoR1
verify using Not1 + Xba1 and BamHI
Result intepretation/Summary
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Conclusion
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