Chiba/Lab Notebook
From 2007.igem.org
(Difference between revisions)
Line 4: | Line 4: | ||
*LINKER LIGATION | *LINKER LIGATION | ||
**pUC19-FliC(vector)の制作 | **pUC19-FliC(vector)の制作 | ||
- | # | + | #pUC19-FliCをInverse PCR(五種類のF-Primer,R-Primer→15通りのPCR) |
##Check Size | ##Check Size | ||
##Digestion(EcoRI,NcoI) | ##Digestion(EcoRI,NcoI) | ||
##Zymo Clean(40μ溶出)→SAP(50μ系)→Zymo Clean(30μ溶出) | ##Zymo Clean(40μ溶出)→SAP(50μ系)→Zymo Clean(30μ溶出) | ||
- | + | #Histag,Histag-antiをアニール | |
- | + | ##Digestion(EcoRI,NcoI) | |
- | # | + | #Ligation |
- | ##Digestion | + | #Transformation |
- | # | + | #miniprep |
- | # | + | #Digestion test |
- | # | + | #Sequence→FliCに6×His配列が挿入されていることを確認 |
- | # | + | |
- | # | + |
Revision as of 05:23, 2 September 2007
Lux TEAM
FliC TEAM
- LINKER LIGATION
- pUC19-FliC(vector)の制作
- pUC19-FliCをInverse PCR(五種類のF-Primer,R-Primer→15通りのPCR)
- Check Size
- Digestion(EcoRI,NcoI)
- Zymo Clean(40μ溶出)→SAP(50μ系)→Zymo Clean(30μ溶出)
- Histag,Histag-antiをアニール
- Digestion(EcoRI,NcoI)
- Ligation
- Transformation
- miniprep
- Digestion test
- Sequence→FliCに6×His配列が挿入されていることを確認