1264-pCMV5-Fc(3c)-NPR1
Cloning of 1264-pCMV5-Fc(3c)-NPR1
[edit]Purpose: Cloning of 1264-pCMV5-Fc(3c)-NPR1
Date: 2012-7-12---2012-7-12
Notebook:0
Page:
Clone ID:1264
Experimenters: yingying
Results
[edit]cloning
[edit]
PCR:
Template: kinase7901 B07
NPR1-Nhe1-F
catgac GCTAGC ATGCCGGGGCCCCGGCGCCCCGCTG
NPR1-Xho1-R wrong primer
gctgacCTCGAGCTAGCCTCGGGTGCTACTCCCCCTC
| band 1 |
band 2 | |
| enzymes |
| |
Colony Screening
[edit]
Result intepretation/Summary
[edit]- <br>
Conclusion
[edit]
Cloning of 1264-pCMV5-Fc(3c)-NPR1
[edit]Purpose: Cloning of 1264-pCMV5-Fc(3c)-NPR1
Date: 2012-7-27---2102-7-30
Notebook:0
Page:
Clone ID:1264
Experimenters: yingying
Results
[edit]cloning
[edit]
PCR
Template:Kinase 7901 B07
primer:NPR1-Nhe1-F+NPR1-Xba1-R
| band 3 |
band 7 | |
| PCRproduct |
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>
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<br>
Cloning of 1264-pCMV5-Fc(3c)-NPR1
[edit]Purpose: Cloning of 1264-pCMV5-Fc(3c)-NPR1
Date: 2012-7-31---2012-8-2
Notebook:0
Page:
Clone ID:1264
Experimenters: yingying
Results
[edit]cloning
[edit]
NPR1-Nhe1-F
catgac GCTAGC ATGCCGGGGCCCCGGCGCCCCGCTG
NPR1-Xba1-R
| band 1 |
band 2 | |
| PCRproduct |
1262,pCMV5-Fc(3C) | |
| enzymes |
Nhe1+Xba1 | |
Colony Screening
[edit]Date: 2012-3-25
Make miniprep and digest with Nco1 + Xba1.
not the right size
Result intepretation/Summary
[edit]- <br>
Conclusion
[edit]<br>
<br>
<br>
Cloning of 1264-pCMV5-Fc(3c)-NPR1
[edit]Purpose: Cloning of 1264-pCMV5-Fc(3c)-NPR1
Date: 2012-8-3--2012-8-6
Notebook:0
Page:
Clone ID:1264
Experimenters: yingying
Results
[edit]cloning
[edit]
| band |
band | |
| PCR product |
1263-2,pCMV5-Fc(3c)-RIPK3 | |
| enzymes |
Nhe+Xba1 | |
Colony Screening
[edit]Date: 2012-8-6
Make miniprep and digest with Nhe+Xba1
NO insert
c
Result intepretation/Summary
[edit]- <br>
Conclusion
[edit]<br>
<br>
<br>
Cloning of 1264-pCMV5-Fc(3c)-NPR1
[edit]Purpose: Cloning of 1264-pCMV5-Fc(3c)-NPR1
Date: 2012-8-7--2012-8-10
Notebook:0
Page:
Clone ID:1264
Experimenters: yingying
Results
[edit]cloning
[edit]
NPR1-Nhe1-F
catgac GCTAGC ATGCCGGGGCCCCGGCGCCCCGCTG
NPR1-Not1-R
| band |
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
sequencing show mis priming
Result intepretation/Summary
[edit]- <br>
Conclusion
[edit]<br>
<br>
<br>
Cloning of 1264-pCMV5-Fc(3c)-NPR1
[edit]Purpose: Cloning of 1264-pCMV5-Fc(3c)-NPR1
Date: 2012-8-16--2012-8-20
Notebook:0
Page:
Clone ID:1264
Experimenters: yingying
Results
[edit]cloning
[edit]
NPR1-Nhe1-F2
catgac GCTAGC ATGCCGGGGCCCCGGCGCCCCGCTG
NPR1-Not1-R
| band 3 |
band 4 | |
| PCR product |
1263 pCMV5-Fc(3c)-RIPK3 | |
| enzymes |
Nhe1+Not1 | |
Colony Screening
[edit]Date: 2012-8-20
Make miniprep and digest with Nhe1+Not1
clone 1,2,3,5,6 may be right
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>
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