The PCR mixture can be divided into several mixture:
1. Using PCR mastermix (1 sample = 50 microliter)
a. water (dH2O) = 10 microliter
b. Primer forward = 5 microliter
c. Primer reverse = 5 microliter
d. Mastermix = 25 microliter
e. DNA sample = 5 microliter
2. Using manual PCR mixture (1 sample = 25 microliter)
a. Buffer polymerase (10x) = 5.5 microliter
b. Magnesium Chloride (25mM) = 2.5 microliter
c. dNTPs = 1 microliter
d. Primer forward = 2.5 microliter
e. Primer reverse = 2.5 microliter
f. Taq polymerase = 0.2 microliter
g. water dH2O = 5.8 microliter
h. DNA sample = 5 microliter
Add-in BSA if MUST to enhance PCR product if the DNA quality is poor.