Comparison of three PCR-based assays for SNP genotyping in plants

Table 2 Thermal cycling parameters for each of the three real-time PCR protocols

	Pre-read stage	Hold stage	1st PCR stage		2nd PCR stage		Post-read stage
	°C/Time	°C/Time	°C/Time	Cycles	°C/Time	Cycles	°C/Time
TaqMan	60 °C/30 s	95 °C/10 min	95 °C/15 s	40			60 °C/30 s
TaqMan	60 °C/30 s	95 °C/10 min	60 °C/1 min	40			60 °C/30 s
KASP	30 °C/1 min	94 °C/15 min	94 °C/20 s	10	94 °C/20 s	29	30 °C/1 min
KASP	30 °C/1 min	94 °C/15 min	61–55 °C/1 min (dropping 0.6 °C per cycle)	10	55 °C/1 min	29	30 °C/1 min
rhAmp	60 °C/30 s	95 °C/10 min	95 °C/10 s	37			60 °C/30 s
			60 °C/30 s
			68 °C/20 s

ISSN: 1746-4811