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	**Ligase: 1ul		**Ligase: 1ul
	**Final volume: 10ul		**Final volume: 10ul
		+	**RT 3h
	*Enzyme Digestion		*Enzyme Digestion
	**pOmpC: 14ul EYFP: 3ul		**pOmpC: 14ul EYFP: 3ul

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Revision as of 09:23, 27 August 2007

• plasmid concentration after liquid culture extraction

part	concentration (ug/ul)
pOmpC	6
EYFP	30
ECFP	23
CinR	15
HSL	49
pCinHSL	7
pCinHSLRBS	12
RBS	14
D-term	27

• Ligation
  • Insert:vector=3:1
  • Vector: 300ng/ul(2ul pOmpC after enzyme digestion)
  • Insert: 100ng/ul(2ul EYFP after enzyme digestion)
  • Ligase: 1ul
  • Final volume: 10ul
  • RT 3h
• Enzyme Digestion
  • pOmpC: 14ul EYFP: 3ul
  • RE: 1ul
  • Heat inactivation: 65°C 20min.
• Culture tar-EnvZ, pLacIQ in 5 ml LB liquid medium (Kan:20ug/ml)