Korea U Seoul/17 September 2010

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(New page: ===Digestion=== # Digestion of PCR products and pSB1A2 ## PyodA-mAAA : EcoRI and SpeI ## PzntA-RFP : XbaI and PstI ## pSB1A3 : EcoRI and PstI <br> {| style="border: 1px solid black;" |- |'...)
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Revision as of 13:22, 20 October 2010

Digestion

  1. Digestion of PCR products and pSB1A2
    1. PyodA-mAAA : EcoRI and SpeI
    2. PzntA-RFP : XbaI and PstI
    3. pSB1A3 : 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
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 4)
  2. Quantification of DNA concentration by NanoDrop
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