From 2007.igem.org

• 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

---

first assembly

• 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

Others

• Culture tar-EnvZ, pLacIQ in 5 ml LB liquid medium (Kan:20ug/ml)