454-pAd5.CMV-cre-iRFP
Cloning of pAd5-CMV-cre-iRFP
Purpose: to clone pAd5-CMV-iRFP by gene synthesis to 418 pAd5.CMV-iRFP
Date: 2011-11-18
Notebook:Lu yingying
Page:
Clone ID:454
Experimenters: lu yingying
Results
PCR Cre with Cre-EcoR1-F + Cre-HindIII-BglII-R, using pTriEx-HTNCre(298) as template
PCR product digest with EcoR1 + Bgl11,
pAd5.CMV-iRFP 418 vector digest with EcoR1 + BamH1
band 1 show insert,band 2 show pAd5.CMV-iRFP 418 vector
digest test:EcoR1+Xho1
Date: 2011-11-22
Notebook:Lu yingying
Page:
Clone ID:454
Experimenters: lu yingying
Results
PCR product digest with EcoR1 + Bgl11, pAd5.CMV-iRFP 418 vector digest with EcoR1 + BamH1
insert and vector
digest with EcoR1 +Xho1
Cloning of pAd5.CMV-cre-iRFP
[edit]Purpose: Cloning of pAd5.CMV-cre-iRFP
Date: 2011-12-14--
Notebook:
Page:
Clone ID:454
Experimenters: yingying
Results
[edit]Subcloning
[edit]Date: 2011-12-14
| 454 |
band1 |
band2 |
| PCR product |
pAd5.CMV-iRFP 418 | |
| digest |
EcoR1 + Bgl11 |
EcoR1 + BamH1 |
Colony Screening
[edit]Date: 2011-12-16
Make miniprep and digest with EcoR1 +Xho1
Lane 1: may right ,need to sequence
Transfect into 293T cells ,then read fluorescence :Ex675nm,Em740nm
| Temperature(°C) |
blank |
blank |
454 |
454 | |
| read14 |
24.7 |
0.501 |
0.483 |
1.065 |
4.195 |
| read30 |
24.9 |
1.743 |
1.322 |
2.207 |
2.343 |
| read54 |
24.8 |
0.486 |
0.899 |
0.958 |
0.898 |
Result intepretation/Summary
[edit]- Sequencing Result:
Conclusion
[edit]Restriction digestion
--Yingying.lu 08:54, 19 December 2011 (EST)