Prepare a PCR tube.

Thaw the KOD One^TM PCR Master Mix (Blue) from the -20°C fridge.

For a 50μL PCR sample, add the following into a PCR tube (follow the sequence):

A	B
KOD OneTM PCR Master Mix (Blue)	25μL
Each Primer (2 primers in total)	1.5μL
DNA Template	1μL
DI Water	21μL

Vortex the PCR tubes to mix well.

Place the PCR tube into the Thermal Cycler with the following conditions:

**Lid Temperature: 105°C; Volume: 50μL

A	B	C
98°C	10s
68°C	5s	Edit: -1°C per cycle
68°C	1 minute
Go to step 1, repeat the cycle 13 times
98°C	10s
55°C	5s
68°C	1 minute
Go to step 5, repeat the cycle 25 times
68°C	5 minutes
12°C	Infinite Loop (∞)

Upon finishing, cancel the current protocol and take out the PCR sample from the Thermal Cycler. Keep the sample in the 4°C fridge for storage. Otherwise, gel electrophoresis may be carried out to isolate the DNA fragment of interest.