Polymerase Chain Reaction (PCR)

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.