NYMU Taipei/Lab Notes/2007 8 27

From 2007.igem.org

(Difference between revisions)
Line 40: Line 40:
*Enzyme Digestion
*Enzyme Digestion
**pOmpC: 14ul EYFP: 3ul
**pOmpC: 14ul EYFP: 3ul
 +
**RE: 1ul
**Heat inactivation: 65°C 20min.
**Heat inactivation: 65°C 20min.
*Culture tar-EnvZ, pLacIQ in 5 ml LB liquid medium (Kan:20ug/ml)
*Culture tar-EnvZ, pLacIQ in 5 ml LB liquid medium (Kan:20ug/ml)

Revision as of 09:21, 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
  • 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)