NYMU Taipei/Lab Notes/2007 8 30

From 2007.igem.org

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== 2nd phase assembly==
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* RBS + cinR
 +
* RBS + HSL
*Enzyme Digestion (RBS)
*Enzyme Digestion (RBS)
**RBS: 11ul
**RBS: 11ul

Revision as of 06:59, 30 August 2007

2nd phase assembly

  • RBS + cinR
  • RBS + HSL
  • Enzyme Digestion (RBS)
    • RBS: 11ul
    • SpeI: 1ul
    • PstI: 1ul
    • 10x buffer: 2, 2ul
    • 10x BSA: 2ul
    • H2O: 3ul
    • Total: 20ul
    • 37°C, 2h
    • Heat inactivation: 65°C 20min
  • Enzyme Digestion (cinR)
    • cinR: 11ul
    • XbaI: 1ul
    • PstI: 1ul
    • 10x buffer: 3, 2ul
    • 10x BSA: 2ul
    • H2O: 3ul
    • Total: 20ul
    • 37°C, 2h
    • Heat inactivation: 65°C 20min
  • Enzyme Digestion (HSL)
    • HSL: 4ul
    • XbaI: 1ul
    • PstI: 1ul
    • 10x buffer: 3, 2ul
    • 10x BSA: 2ul
    • H2O: 10ul
    • Total: 20ul
    • 37°C, 2h
    • Heat inactivation: 65°C 20min
  • Plasmid extraction: H_IGFBP3, H_IGFBP7, H_INS, M_IDE
    • From 5ml LB medium to 50ul EB
  • Ligation (RBS+cinR)
    • RBS: 1ul
    • cinR: 1ul
    • 10x buffer: 1ul
    • T4 ligase (Promega): 1ul
    • H2O: 6ul
  • Ligation (RBS+HSL)
    • RBS: 1ul
    • HSL: 1ul
    • 10x buffer: 1ul
    • T4 ligase (Promega): 1ul
    • H2O: 6ul