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. 2024 Oct 30;12(11):2189. doi: 10.3390/microorganisms12112189

Table 1.

PCR amplification system and conditions.

Gene Segment Primer Reaction System Reaction Conditions
16S rDNA 27F(5′-AGAGTTTGATCCTGGCTCAG-3′)
1492R(5′-GGTTACCTTGTTACGACTT-3′)		 Mix 12.5 μL
primer 0.5 μL
template 1.0 μL
ddH2O 11.0 μL		 95 °C pre-denaturation 5 min. 95 °C degeneration 30 s, 58 °C annealing 15 s, 72 °C elongation 2 min, 20 cycles. 72 °C elongation 10 min.
gyrB UP-1(5′-GAAGTCATCATGACCGTTCTGCAYGCNGGNGGNAARTTYGA-3′)
UP-2r(5′-AGCAGGGTACGGATGTGCGAGCCRTCNACRTCNGCRTCNGTCAT-3′)		 95 °C pre-denaturation 5 min. 94 °C degeneration 1 min, 54 °C annealing 1 min, 72 °C elongation 2 min, 25 cycles. 72 °C elongation 10 min.
rpoB f(5′-AGGTCAACTAGTTCAGTATGGAC-3′)
r(5′-AAGAACCGTAACCGGCAACTT-3′)		 94 °C pre-denaturation 4 min. 94 °C degeneration 1 min, 51 °C annealing 1 min, 72 °C elongation 1 min, 25 cycles. 72 °C elongation 10 min.