TABLE 1.

PCR conditions and primers used for the reactions

Step	PCR program		Primers (amt [pmol])
	Temp (°C), duration	No. of cycles
RT-PCRa (template amplification)	55, 30 min	1	MSRT1 (12), MSRT2 (12)
	95, 2 min	1
	94, 30 s; 58, 30 s; 72, 30 s	7
Second PCRb (detection)	94, 30 s; 58, 30 s; 72, 30 s	20	41 MS-PCR: MSRT1 (3), MS41W (2), MS41M (3)
			70MS-PCR: MSRT1 (5), MS70W (2), MS70M (4)
Third PCRb (detection)	94, 30 s; 58, 30 s; 72, 30 s	40	41MS-PCR: MSRT3 (10), MS41W (8), MS41M (20)
			70MS-PCR: MSRT3 (10), MS70W (8), MS70M (20)

^aRT-PCR: 5 mM MgCl₂, 1 mM deoxynucleoside triphosphates, 1 × One-Step PCR buffer (Takara).

^bSecond and third PCRs 1mM MgCl₂, 0.08 mM deoxynucleoside triphosphates, 1 × PCR buffer (Promega).