1262-pCMV5-Fc(3C)
Cloning of 1262-pCMV5-Fc(3C)
[edit]Purpose: Cloning of 1262-pCMV5-Fc(3C)
Date: 2012-7-6---2012-7-9
Notebook:0
Page:
Clone ID:1262
Experimenters: yingying
Results
[edit]subcloning
[edit]
| band 5 |
band 6 | |
| pLex-CMV5-CuO 448 |
1219 pCDNA3.3-Fc(3C) | |
| enzymes |
Nde1 + BamH1 | |
1219: pCDNA3.3-Fc(3C), Nde1+ BamH1 should digest out 417 bp insert (CMV promoter, from 184 to 901). If there are three bands, it is unlikely correct.
448: pLex-CMV5-CuO, Has two Nde1 sites (2964, 7933), BamH1 is 3979, CMV5 promoter is 1015 bp
Colony Screening
[edit]Date: 2012-7-9
Make miniprep and digest with Nde1 + BamH1
clone 1,2,3,4,5,6 right
Need to sequence 1262, it is unlikely good.
Result intepretation/Summary
[edit]
Conclusion
[edit]
Cloning of 1262-pCMV5-Fc(3C)
[edit]Purpose: Cloning of 1262-pCMV5-Fc(3C)
Date: 2012-7-17---2012-7-19
Notebook:0
Page:
Clone ID:1262
Experimenters: yingying
Results
[edit]subcloning
[edit]
| insert |
lane 1 | |
| pLex-CMV5-CuO 448 |
1219 pCDNA3.3-Fc(3C) | |
| enzymes |
Nde1 + BamH1 | |
Colony Screening
[edit]Date: 2012-7-19
Make miniprep and digest with Nde1 + BamH1
clone 1,2,3,5,6,7,8 may be right
digest detect again with Xho1
8/2/2012; Digest 1262 with Xba1, BamH1, Nhe1
| 1 | 2 | 3 | 4 |
| Xba1+ BamH1 | Xba1 | Nhe1 | Nhe1+ BamH1 |
Is it possible there are two Xba1 sites? or star activity/overdigestion?
Result intepretation/Summary
[edit]- <br>
Conclusion
[edit]<br>
<br>
<br>