PCR 2X Master Mix

The master mix can be purchased premixed commercially or mixed from the individual components.

For a single 25 µl reaction

2.5 µl 10X Taq Buffer (usually this is supplied with the polymerase, this assumes it does not already have MgCl₂.)

0.5 µl 10mM dNTPs

1 µl 50mM MgCl₂ (adjusting the Mg concentration affects the specificity of the polymerase and can be optimized)

0.5 µl (5 U / µl) thermostable (Taq) polymerase

8 µl nuclease-free H₂O (adjust if necessary if MgCl₂ or Taq volume is adjusted for a 12.5 µl total volume).

In general a mix would be made for many reactions, say for 100, and the volumes multiplied accordingly. The mix can be stored at -20 C.

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