306 pAAV-EF1-MCS
Cloning of EF to pAAV-CMV-MCS
[edit]Purpose: Cloning of EF to pAAV-CMV-MCS.
Date: 2011-8-8
Notebook:
Page:
Clone ID:
Experimenters: Haiyan
Results
[edit]Subcloning
[edit]Date: 2011-6-8
Colony Screening
[edit]Date: 2011-8-8
Make miniprep and digest with Mlui+ecori
Lane 1: marker
Lane 2-5: 295_pAAV-Syn-MCS, Mlui+ecori
Lane 6-8: 306_pAAV-EF-MCS, Mlui+ecori
Looks 295 no positive clone, 306 looks have insert, need to do pcr additional confirm.
Result intepretation/Summary
[edit]- PKAalpha 3+4 has one internal Xho1 site. GFP-Puro can be cut out with Xho1 + Mlu1. So Lane 1 has correct number of bands.
- PKA alpha 3+4 with CMV promoter can be cut out with Nde1+ BamH1. Lane 5 has correct number of bands.
- lane 6 maybe good (incomplete digestion)
- Sequencing Result:
Conclusion
[edit]Restriction digestion
shows pLentiGP clone #1 may be good,
pLVGP-PKAalpha 3+4 is good
[edit]
Paul.xu 08:23, 15 June 2011 (EST)