Master mix (90 µl) contents were 44 µl

DEPC treated H2O, 20 µl 5×buffer, 6 µl MgCl2, 16 µl d NTP’s (1.25 mµ), 2

µl Forward primer and 2 µl Reverse primer.

Test sample (45 µl of Master Mix) added

with 5 µl of c DNA and 0.3 µl G. Taq Polymerase. Control (45 µl of Master Mix) was added with 5 µl of DEPC+ H2O and 0.3 µl G.Taq Polymerase.

Proceed to Thermocycler.

This might help you.