NYMU Taipei/Lab Notes/2007 8 27

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(Difference between revisions)
Line 3: Line 3:
!part
!part
!concentration (ug/ul)
!concentration (ug/ul)
 +
!A260 (OD)
|-
|-
|pOmpC
|pOmpC
|6
|6
 +
|0.006
|-
|-
|EYFP
|EYFP
|30
|30
 +
|0.030
|-
|-
|ECFP
|ECFP
|23
|23
 +
|0.023
|-
|-
|CinR
|CinR
|15
|15
 +
|0.015
|-
|-
|HSL
|HSL
|49
|49
 +
|0.049
|-
|-
|pCinHSL
|pCinHSL
|7
|7
 +
|0.007
|-
|-
|pCinHSLRBS
|pCinHSLRBS
|12
|12
 +
|0.012
|-
|-
|RBS
|RBS
|14
|14
 +
|0.014
|-
|-
|D-term
|D-term
|27
|27
 +
|0.027
|-  
|-  
|}
|}

Revision as of 14:54, 11 September 2007

  • plasmid concentration after extraction
part concentration (ug/ul) A260 (OD)
pOmpC 6 0.006
EYFP 30 0.030
ECFP 23 0.023
CinR 15 0.015
HSL 49 0.049
pCinHSL 7 0.007
pCinHSLRBS 12 0.012
RBS 14 0.014
D-term 27 0.027

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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