1234 pET30b-M1wt-1-165
Cloning of 1234 pET30b-M1wt-1-165
[edit]Purpose: Cloning of ORF for 1-165 aa of wt Flu M1
Date: 2012-6-5
Notebook:
Page:
Clone ID: 1204
Experimenters: Paul
Results
[edit]cloning
[edit]Date: 5-29-2012
PCR using Kapa HiFi.
cut out band and purify using gel extraction kit. digest with Nde1 + Xho1. ligate into Nde1 + Xho1 digested pET28b.
transform DH5alpha
Colony Screening
[edit]Date: 6/7/12
digest miniprep with Nde1 + Xho1
send clone #6 to sequencing
sequecing: #6: no priming, #1 is MMLV (contamination)
6/10/2012
redo PCR:
6/11/2012
PCR
1, 2: mutant
3,4: wt
Digest vector and PCR product with Nde1 + Xho1
1: pET30b
2: PCR product
digest miniprep with Nde1 + Xho1
2012-6-20
digest detect :Nde1+Xho1
Result intepretation/Summary
[edit]
Conclusion
[edit]Cloning of 1234 pET30b-M1wt-1-165
[edit]Purpose: Cloning of ORF for 1-165 aa of wt Flu M1
Date: 2013-7-17---2013-7-18
Notebook: 006026
Page:006209
Clone ID: 1234
Experimenters: Paul
Results
[edit]cloning
[edit]template is wt M1
FluASeg-7-NdeI-F
FluASeg-7-Xho1-R
| lane |
1 |
2 |
3 |
| PCR product |
pET28b |
501,pET30b-MMLV | |
| enzymes |
Nde1+Xho1 | ||
screening
[edit]