PCR Component

Concentration

Volume (µL)

DNA template

-

4.0 - 5.0

ddH₂O

-

10 - 14

Buffer

5 X

2.5

MgCl₂

25 mM

3 - 4

Mixture of dNTP

1 mM

0.5

Primer forward

15 - 30 pmol∙µL⁻¹

0.5

Primer reverse

15 - 30 pmol∙µL⁻¹

0.5

Tag DNA polymerase

4 - 6 U∙µL⁻¹

0.3