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	== FliC TEAM ==		== FliC TEAM ==
-	*LINKER LIGATION	+	*pUC19-FliC-Hisの作製
-	**pUC19-FliC(vector)の制作	+	**LINKER LIGATION
	#pUC19-FliCをInverse PCR（五種類のF-Primer,R-Primer→15通りのPCR）		#pUC19-FliCをInverse PCR（五種類のF-Primer,R-Primer→15通りのPCR）
	##Check Size		##Check Size

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Revision as of 05:25, 2 September 2007

Lux TEAM

FliC TEAM

• pUC19-FliC-Hisの作製
  • LINKER LIGATION

1. pUC19-FliCをInverse PCR（五種類のF-Primer,R-Primer→15通りのPCR）
   1. Check Size
   2. Digestion(EcoRI,NcoI)
   3. Zymo Clean(40μ溶出)→SAP（50μ系）→Zymo Clean（30μ溶出）
2. Histag,Histag-antiをアニール
   1. Digestion(EcoRI,NcoI)
3. Ligation
4. Transformation
5. miniprep
6. Digestion test
7. Sequence→FliCに6×His配列が挿入されていることを確認