Table 4.

Composition of the PCR mixture and temperature profile for the bacterial gene 16S rRNA amplification

Real Time 2x-PCR Mix SYBR A	10 μl
Starter 1055F 10 μM 5′-ATGGCTGTCGTCAGCT-3’	0.4 μl
Starter 1392R 10 μM 5′-ACGGGCGGTGTGTAC-3’	0.4 μl
DNA template	1 μl
deionized water	8.2 μl
initial denaturation	95 °C, 3 min.
denaturation	95 °C, 15 s
priming	58 °C, 30 s
elongation	72 °C, 30s
appointment of the melting curve (measurement of the fluorescence in each temperature)	65 °C - > 95 °C

Abbreviations: PCR Polymerase chain reaction, PCR Mix SYBR A – ready-to-use mixture to the real-time PCR with fluorophore (A&A Biotechnology, Poland); F = forward primer; R = reverse primer