Direct PCR from muscle tissue of different animals

A 237 bp DNA fragement was amplified with Phire® Hot Start DNA Polymerase directly from muscle tissue of various animals. 0.4 mg of muscle tissue was added directly into a 20 µl PCR reaction. Piko® Thermal Cycler and UTW® reaction vessels were used in PCR.

The following 2-step protocol was used:
98°C for 5 min, 40 cycles of [98°C for 5 s, 72°C for 20 s], 72°C for 1 min.

Comparing performance of Phire^® DNA Polymerase to Taq DNA polymerase

Two different DNA fragments were amplified either with Phire Hot Start DNA Polymerase or a hot start Taq DNA polymerase from differently prepared templates. Piko Thermal Cycler and UTW reaction vessels were used in PCR.

The following PCR protocols were used:
Phire Hot Start DNA Polymerase : 98°C for 5 min, 40 cycles of [98°C for 5 s, 60°C for 20 s, 72°C for 20 s], 72°C for 1 min.
Taq DNA polymerase : 95°C for 5 min, 40 cycles of [94°C for 30 s, 60°C for 30 s, 72°C for 1 min], 72°C for 10 min.

1: DNA extracted with a commercial kit
2: 0.4 mg of bear muscle tissue directly added into the reaction
3: 0.4 mg of bear muscle tissue was first incubated in 50 µl TE buffer at 50°C for 3 min. 1 µl of the supernatant was used in the PCR reaction.

General notes