1336 pCMV5-Fc(3C)-Dest-WPRE
Cloning of 1336 pCMV5-Fc(3C)-Dest-WPRE
[edit]Purpose: Cloning of WPRE into vector pCMV5-Fc(3C)-Dest to make 1336 pCMV5-Fc(3C)-Dest-WPRE
Date: 2012-8-17 to
Notebook:
Page:
Clone ID: 1328
Experimenters: yingying
Results
[edit]cloning
[edit]Date:2012-8-17
PCR WPRE from 90 pLVX-puro-HA-AKR1C4
WPRE-Not1-F atcgatGCGGCCGC AATCAACCTCTGGATTACAAAATTTG
WPRE-Xba1-R ttagatTCTAGACCAGGCGGGGAGGCGGCCCAAAGGGAGAT
Cut out band, purify and digest with Not1 + Xba1
digest vector 1328 pCMV5-Fc(3C)-Dest with Not1 + Xba1, ligate and transform ccdB survival
Colony Screening
[edit]Date:
Gateway reaction
[edit]
Result intepretation/Summary
[edit]Conclusion
[edit]Cloning of 1336 pCMV5-Fc(3C)-Dest-WPRE
[edit]Purpose: Cloning of WPRE into vector pCMV5-Fc(3C)-Dest to make 1336 pCMV5-Fc(3C)-Dest-WPRE
Date: 2012-8-20 to
Notebook:
Page:
Clone ID: 1336
Experimenters: yingying
Results
[edit]cloning
[edit]Date:2012-8-20
PCR WPRE from 90 pLVX-puro-HA-AKR1C4
WPRE-Not1-F atcgatGCGGCCGC AATCAACCTCTGGATTACAAAATTTG
WPRE-Xba1-R ttagatTCTAGACCAGGCGGGGAGGCGGCCCAAAGGGAGAT
Cut out band, purify and digest with Not1 + Xba1
digest vector 1328 pCMV5-Fc(3C)-Dest with Not1 + Xba1, ligate and transform ccdB survival
Colony Screening
[edit]Date: 2012-8-22
digest : Not1 + Xba1
clone 3,4 may be right
Gateway reaction
[edit]2012-8-29
miniprep and digest with Kpn1+Xho1
1--8:393+1336-3/4; 9:1336-3/4
need to transfect to test EGFP expression