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In: Biology

You have the following stock solutions available to prepare your PCR master mix. 250mM Tris-HCl stock...

  1. You have the following stock solutions available to prepare your PCR master mix.

250mM Tris-HCl stock solution

400mM KCl stock solution

20mM MgCl2 stock solution

15mM dNTP stock solution

10uM Forward primer stock solution

10uM Reverse primer stock solution

10 U/uL Taq polymerase

4 ng/uL Template DNA

Perform the necessary calculations to determine the volume of each component needed to achieve the below specified concentrations per reaction. When performing your calculations, be sure that you will not be required to pipette a volume less than 0.5uL, for the greatest accuracy. Use nuclease-free water to bring the total reaction volume to 25uL.

  • 30mM Tris-HCl
  • 40mM KCl
  • 1.5mM MgCl2
  • 0.2mM dNTPs
  • 200nM Forward primer
  • 200nM Reverse primer
  • 3U Taq polymerase
  • 1ng Template DNA

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