From 2014.igem.org

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25^th August

• Chewback : miniprep


• Assembly : ELP-ELP, ELP-RLP, ELP-SLP, SLP-RLP, SLP-SLP and RLP-RLP
  
  • Digestion
    All OK but try again the plasmid pSB1C3 digestion
    
    incubation 1h30 at 37°C :
    

    Enzyme 1 (Nhe1 or X)	0,5μl
    Enzyme 2 (S)	0,5lμl
    10X Buffer 4	5μl
    DNA	10μl
    H2O	qsp 50μl

    - Enzyme inactivated 20min at 80°C
    - Gel 1% (30min, 100V)
  
  
  • Ligation

    Plasmid	4μl
    Insert	4μl
    Buffer 10X	1μl
    Taq ligase	0,5μl
    H2O	qsp 10μl

    30min at 45°C°C
  
  
  • Transformation
  
  
  
  
  
  Purification
  
  
  • Dialysis at 11h
  • Lyophilisation at 12h10