Add the primers, the DNA template, and the following reagents from the into a PCR tube to make a 50µL PCR sample:

A	B
10x Mutazyme II (10x Reaction Buffer)	5μL
40mM dNTP Mix	1μL
DI water	41.5μL
Each Primer (both forward & reverse)	0.5μL
Mutazyme II	1μL
DNA Template	0.25μL

Mix the solution well.

Place the sample into the Thermal Cycler and set it with the following conditions:

A	B	C
Initial Denaturation	95°C	2 minutes
Denaturation	95°C	1 minute
Annealing	55°C	1 minute
Extension	72°C	1 minute
Go to step 2, repeat the cycle 44 times
Extension	72°C	10 minutes
Finish	12°C	Infinite Loop

Upon finishing the PCR steps, add 10µL of DNA loading dye.

Proceeds to the gel electrophoresis to isolate the gene fragment of interest.