- You have the following stock solutions available to prepareyour 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 ofeach component needed to achieve the below specified concentrationsper reaction. When performing your calculations, be sure that youwill not be required to pipette a volume less than 0.5uL, for thegreatest accuracy. Use nuclease-free water to bring the totalreaction 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
