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Revision as of 07:02, 30 August 2007 (view source) Lihsiangyen (Talk | contribs) (→2nd phase assembly) ← Older edit		Revision as of 07:15, 30 August 2007 (view source) Lihsiangyen (Talk | contribs) (→2nd phase assembly) Newer edit →
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	== 2nd phase assembly==		== 2nd phase assembly==
-	* RBS + cinR	+	* '''Objective: RBS + cinR'''
-	* RBS + HSL	+	* '''Objective: RBS + HSL'''
	*Enzyme Digestion (RBS)		*Enzyme Digestion (RBS)
	**RBS: 11ul		**RBS: 11ul

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Revision as of 07:15, 30 August 2007

2nd phase assembly

• Objective: RBS + cinR
• Objective: RBS + HSL
• Enzyme Digestion (RBS)
  • RBS: 11ul
  • SpeI: 1ul
  • PstI: 1ul
  • 10x buffer: 2, 2ul
  • 10x BSA: 2ul
  • H2O: 3ul
  • Total: 20ul
  • 37°C, 2h
  • Heat inactivation: 65°C 20min
• Enzyme Digestion (cinR)
  • cinR: 11ul
  • XbaI: 1ul
  • PstI: 1ul
  • 10x buffer: 3, 2ul
  • 10x BSA: 2ul
  • H2O: 3ul
  • Total: 20ul
  • 37°C, 2h
  • Heat inactivation: 65°C 20min
• Enzyme Digestion (HSL)
  • HSL: 4ul
  • XbaI: 1ul
  • PstI: 1ul
  • 10x buffer: 3, 2ul
  • 10x BSA: 2ul
  • H2O: 10ul
  • Total: 20ul
  • 37°C, 2h
  • Heat inactivation: 65°C 20min
• Plasmid extraction: H_IGFBP3, H_IGFBP7, H_INS, M_IDE
  • From 5ml LB medium to 50ul EB
• Ligation (RBS+cinR)
  • RBS: 1ul
  • cinR: 1ul
  • 10x buffer: 1ul
  • T4 ligase (Promega): 1ul
  • H2O: 6ul
• Ligation (RBS+HSL)
  • RBS: 1ul
  • HSL: 1ul
  • 10x buffer: 1ul
  • T4 ligase (Promega): 1ul
  • H2O: 6ul

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by 立翔