Team:Todai-Tokyo/Protocols/PCR/Enzymes

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Contents

Taq Polymerases

Pfu Ultra

Unless otherwise stated, we use this Taq

  • 0.2µl 100µM 5’primer
  • 0.2µl 100µM 3’primer
  • 1.6µl 2.5mM dNTP
  • 2µl 10 x Pfu Ultra 2 buffer
  • 15.45µl MilliQ
  • 0.05µl DNA (to a final concentration of 5-30ng/20ul)
  • 0.5µl Pfu Ultra2


Ex Taq

  • 0.2µl 100µM 5’primer
  • 0.2µl 100µM 3’primer
  • 1.6µl 2.5mM dNTP
  • 2µl 10 x Pfu Ultra 2 buffer
  • Xµl MilliQ(up to 20ul)
  • Xµl DNA(to a final concentration of 200ng/20ul)
  • 0.2µl Ex-Taq

KOD plus

  • 5μl 10 × PCR Buffer for KOD-Plus
  • 5μl 2mM dNTPs
  • 2μl 25mM MgSO4
  • 1.5μl 10μM primer 1
  • 1.5μl 10μM primer 2
  • >1 μl template DNA
    • Genomic DNA 10~200ng/50μl
    • Plasmid DNA 1~50ng/50μl
  • 1 μl KOD-Plus
  • MilliQ to 50μl