Table 1.

PCR primers and thermal cycling conditions

Methods	Primers	Sequence
BSP		Forward: 5'-AAGAAAAGAAAAGAAAAGAAAATTTAG-3'
		Reverse: 5'-TTTAGTGAGGTAGATAATTTTTT-3'
PCR conditions		Initial denaturation at 95°C (10 min) and 35 cycles at 95°C (3 s), 52°C (3 s), 72°C (5 s), and a final extension at 72°C (10 s)
MSP	Methylation-	Forward: 5'-TAGATAATTTTTTTTAGATTTTGCGGTCG-3'
	Specific	Reverse: 5'-TCCTCGCGAAACTACGC-3'
PCR condition		Initial denaturation at 95°C (10 min) and annealing temperature decrement of 0.5°C every cycle (from 70°C to 66.5°C) followed by 32 cycles of 66°C (3 s), 72°C (5 s), and a final extension at 72°C (10 s)
MSP	Non-methylation	Forward: 5'-TAGATAATTTTTTTTAGATTTTGTGGTTG-3'
	-specific	Reverse: 5'-TCCTCACAAAACTACAC-3'
PCR condition		Initial denaturation at 95°C (10 min) and annealing temperature decrement of 0.5°C every cycle (from 64°C to 60.5°C) followed by 32 cycles of 60°C(3 s), 72°C (5 s), and a final extension at 72°C (10 s)