From 2014.igem.org

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19^th August

• Verification of our plasmid Miniprep  we haven’t any problems with the plasmid
• Digestion of 1μg of insert
  Ex : ELP = 460ng/μl
  
  So 1μg (ELP) = 2,17μl
  + 7,83μl H2O
  
  DNA
  10μl
  
  Xba1
  0,5μl
  
  Spe1
  0,5μl
  
  10X Buffer 4
  5μl
  
  H+O
  qsp 50μl
  
  
  Migration on agarose 1,5% gel (100V, 30min)
  


• Test of pSB1C3 miniprep
  • Digestion : we want 1μg of plamisd
  • Ligation : [insert] = 6 [plasmid] => molar concentration


• PCR: linearization of ELP-40

• Problem with our 1x500ml Erlenmeyer  bacteria grow slower than in 2x250ml erlen.


• Preculture of ELP-60 in 2x250ml of TB+amp


• Preculture of BL21 in 5ml of LB overnight