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

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

Reaction	Final concentration
Components	egc	khe	cps	gyrB	omp P6	copB
TaqMan Universal PCR Mastermix	1×	1×	1×	1×	1×	1×
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

ISSN: 1471-2466