NYMU Taipei/Lab Notes/2007 8 27

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)