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Table 2 Optimized conditions for egc, khe, cps, gyrB, omp P6 and copB RT-qPCR assay

From: Quantitative analysis of pathogens in the lower respiratory tract of patients with chronic obstructive pulmonary disease

Reaction Final concentration
Components egc khe cps gyrB omp P6 copB
TaqMan Universal PCR Mastermix
Primer 0.6 μmol/L 0.6 μmol/L 0.6 μmol/L 0.6 μmol/L 0.6 μmol/L 0.6 μmol/L
Probe 0.3 μmol/L 0.3 μmol/L 0.3 μmol/L 0.3 μmol/L 0.3 μmol/L 0.3 μmol/L
Total volume 20 μl 20 μl 20 μl 20 μl 20 μl 20 μl
Melt 95 °C, 10 min 94 °C, 4 min 94 °C, 4 min 95 °C, 10 min 94 °C, 4 min 94 °C, 4 min
Denaturation 95 °C, 15 s 94 °C, 30 s 94 °C, 30 s 95 °C, 15 s 94 °C, 30 s 94 °C, 30 s
Annealing/Extension 60 °C, 60 s 60 °C, 60 s 60 °C, 60 s 60 °C, 60 s 60 °C, 60 s 60 °C, 60 s
Cycles 45 40 40 45 40 40
PCR Product Size 82 base pairs 77 base pairs 67 base pairs 190 base pairs 156 base pairs 71base pairs