From 2014.igem.org

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12^th August

• Digestion We want a DNA concentration of 100ng/μl
  Time-Saver protocol
  
  DNA 10μl
  Xba1 0,5μl
  Spe1 0,5μl
  Buffer 10X 5μl
  H2O qsp 50μl
  
  => 15min at 37°C
  => Inactivation of enzyme 20min at 80°C

Final concentration after digestion :
• Plasmid: 18 ng/μl
• Insert: 20 ng/μl
• Ligation
  
  ELP – 369bp
  SLP – 369bp
  RLP – 357bp
  Plasmid – 2270bp
  Plasmid 4μl
  Insert 1,8μl
  Buffer 10X 1μl
  Taq ligase 0,5μl
  H2O qsp 10μl
  
• Making competent cells
  BL21 in LB+Amp plates