Team:UT-Tokyo/PCR(KODplus)

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UT-Tokyo

PCR(KODplus)

Preparation

  • Components Volume Final Concentration
  • 10× PCR Buffer for KOD -Plus- 5 μl 1×
  • 2mM dNTPs 5 μl 0.2mM each
  • 25mM MgSO4 2 μl 1.0mM
  • Primer (10μM each) 1.5 μl 0.3μM each
  • Template DNA ≧ 1 μl
  • Genomic DNA 10~200 ng/50μl
  • (Plasmid DNA 1~50 ng/50μl)
  • KOD -Plus- (1U/μl) 1 μl 1 U/50μl
  • MiliQ to 50 μl

PCR program

  • Pre Denature : 94 degrees, 2min.
  • Denature : 94 degrees, 15sec.
  • Annealing : (Tm-5) degrees, 30sec.
  • Extension : 68 degrees, 1min. /kb
  • Set extension time for 1 min./kb