NYMU Taipei/Lab Notes/2007 8 27

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* plasmid concentration after liquid culture extraction
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* plasmid concentration after extraction
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Revision as of 07:13, 30 August 2007

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

Phase 1 assembly

  • Objective: pOmpC + (RBS+EYFP)
  • 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)
    • pOmpC: 14ul
    • SpeI: 1ul
    • PstI: 1ul
    • 10x buffer: 2, 2ul
    • 10x BSA: 2ul
    • Total: 20ul
    • 37°C, 2h
    • Heat inactivation: 65°C 20min
  • Enzyme Digestion (EYFP)
    • EYFP: 3ul
    • XbaI: 1ul
    • PstI: 1ul
    • 10x buffer: 3, 2ul
    • 10x BSA: 2ul
    • H2O: 11ul
    • Total: 20ul
    • 37°C, 2h
    • Heat inactivation: 65°C 20min

Others

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

Who attend?

  • 立翔
  • 德銘
  • 惇茹
  • 香君
  • 宥嫺
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