1269-pCMV5-Fc(3c)-GNE
Cloning of 1269-pCMV5-Fc(3c)-GNE
[edit]Purpose: Cloning of 1269-pCMV5-Fc(3c)-GNE
Date: 2012-7-27---2012-7-30
Notebook:0
Page:
Clone ID:1269
Experimenters: yingying
Results
[edit]cloning
[edit]
PCR using
GNE-Nhe1-F atcgca gct agc ATGGAGAAGAATGGAAATAAC
GNE-Xba1-R gatgctTCTAGACtAGTAGATCCTGCGTGTTGTGTAG
| band 3 |
band 7 | |
| PCR product |
1262,pCMV5-Fc(3C) | |
| enzymes |
Nhe1 + Xba1 | |
Colony Screening
[edit]Date: 2012-7-30
Make miniprep and digest with Nhe1 + Xba1
no insert
Result intepretation/Summary
[edit]- <br>
Conclusion
[edit]<br>
<br>
<br>
Cloning of 1269-pCMV5-Fc(3c)-GNE
[edit]Purpose: Cloning of 1269-pCMV5-Fc(3c)-GNE
Date: 2012-7-31---2012-8-2
Notebook:0
Page:
Clone ID:1269
Experimenters: yingying
Results
[edit]cloning
[edit]
GNE-Nhe1-F
GNE-Xba1-R
| band 5 |
band 6 | |
| PCR product |
1262,pCMV5-Fc(3C) | |
| enzymes |
Nhe1 + Xba1 | |
Colony Screening
[edit]Date: 2012-8-2
Make miniprep and digest with Nhe1 + Xba1
no right clones
Result intepretation/Summary
[edit]- <br>
Conclusion
[edit]<br>
<br>
<br>
Cloning of 1269-pCMV5-Fc(3c)-GNE
[edit]Purpose: Cloning of 1269-pCMV5-Fc(3c)-GNE
Date: 2012-8-3---2012-8-6
Notebook:0
Page:
Clone ID:1269
Experimenters: yingying
Results
[edit]cloning
[edit]
| band 1 |
band 2 | |
| PCR product |
1263-2pCMV5-Fc(3c)-RIPK3 | |
| enzymes |
Nhe1+Xba1 | |
Colony Screening
[edit]Date: 2012-8-6
Make miniprep and digest with Nhe1+Xba1
No insert
c
Result intepretation/Summary
[edit]- <br>
Conclusion
[edit]<br>
<br>
<br>
Cloning of 1269-pCMV5-Fc(3c)-GNE
[edit]Purpose: Cloning of 1269-pCMV5-Fc(3c)-GNE
Date: 2012-8-7---2012-8-10
Notebook:0
Page:
Clone ID:1269
Experimenters: yingying
Results
[edit]cloning
[edit]
GNE-Nhe1-F
GNE-Not1-R
| band 1 |
band | |
| PCR product |
1263,pCMV5-Fc(3c)-RIPK3 | |
| enzymes |
Nhe1+Not1 | |
Colony Screening
[edit]Date: 2012-8-10
Make miniprep and digest with Nhe1+Not1
9/3/2012
Digest Maxiprep DNA with
Nhe1+ Not1--> GOI
Not1 + Xba1--> to see if SVLT is in the vector
| 1 |
2 |
3 |
4 |
5 |
6 |
7 |
8 |
9 |
10 |
11 |
12 |
| 1224 |
1263 |
1264 |
1269 |
1276 |
1284 |
1224 |
1263 |
1264 |
1269 |
1276 |
1284 |
| Nhe1+ Not1 |
Nhe1+ Not1 |
Nhe1+ Not1 |
Nhe1+ Not1 |
Nhe1+ Not1 |
Nhe1+ Not1 |
Not1+ Xba1 |
Not1+ Xba1 |
Not1+ Xba1 |
Not1+ Xba1 |
Not1+ Xba1 |
Not1+ Xba1 |
| lot of RNA | lot of RNA |
1224: vector with SVLT, digest with Not1 + Nhe1: why so many bands??
1263: why multiple bands with Nhe1 + Not1 digestion? there is an internal Xba1 site, digest with
Not1 + Xba1 --> 500 bp
Transfection: cell aggreggation: 1284 and 1276
Result intepretation/Summary
[edit]- <br>
Conclusion
[edit]<br>
<br>
<br>