PAAV-cre 256
SubCloning of Cre to pAAV-MCS
[edit]Purpose: Subcloning of Cre to pAAV-MCS
Date: 2011-7-? to 2011-7-?
Notebook:
Page:
Clone ID:
Experimenters: Haiyan/Paul
Results
[edit]Subcloning
[edit]Date: 2011-6-8
Digest pLVGP-cre (clone #236) with Ecori + XhoI
Digest pAAV-MCS (clone #241) with Ecori + XhoI
cut out insert and vector, ligate and transform STBL3
Lane 1: Marker
Lane 2: pLVGP-cre , EcoR1 + XhoI
Lane 3: pAAV-MCS, EcoRI + XhoI
Colony Screening
[edit]Digestion
Lane 1: pAAV-Cre #1, digest with EcoR1 + Xho1
Lane 2: pAAV-Cre #2, digest with EcoR1 + Xho1
Lane 3: pAAV-Cre #3, digest with EcoR1 + Xho1
Lane 4: pDs-AAV-SMN1 #1, digest with BglII + Pst1
Lane 5: pDs-AAV-SMN1 #2, digest with BglII + Pst1
Lane 6: pDs-AAV-SMN1 #3, digest with BglII + Pst1
Lane 7: pAAV-Cre #1, digest with Nco1 + Xho1
Lane 8: pAAV-Cre #2, digest with Nco1 + Xho1
Lane 9: pAAV-Cre #3, digest with Nco1 + Xho, well leaks
Lane 10: pET30b , digest with Nde1 + Nco1
--Paul.xu 12:15, 17 July 2011 (EST)
Sequencing
Result intepretation/Summary
[edit]Looks like pAAV-Cre #2 & #3 are good. There is one Nco1 site in the CMV promoter
Conclusion
[edit]