Team:Todai-Tokyo/Protocols/PCR/Enzymes

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Contents

Taq Polymerases

Pfu Ultra

Unless otherwise stated, we use this polymerase

  • 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
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