Korea U Seoul/1 October 2010

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Digestion



  1. Check : recombinant plasmid sequence
  2. Selection of correct clones
  3. Digestion of PCR products(ParsR-GFP) and pSB1C3
    1. ParsR-GFP : EcoRI and SpeI
    2. pSB1C3 : EcoRI and PstI


Reagent Volume (μL)
DNA (about 30ng/μL) 30
10x NEB buffer 2 5
BSA (10mg/mL) 0.5
Appropriate 1st and 2nd restriction enzymes 2 (each 1)
D.W. 12.5 / total = 50
colspan="2" Completely digestion at 37˚C for 2 hours (at least)


and stop at 80˚C for 20min


  1. Confirmation of digested products by agarose gel electrophoresis (Figure 6)
  2. Quantification of DNA concentration by NanoDrop
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