Table 3.

List of primers and probe assayed in RT-PCR reactions

RT-PCR MiX	Forward primer at final concentration of:	Reverse primers at final concentration of:	Probe at final concentration of:
#1 primers from Grywna (2010)	F1, 500 nM	R1, 500 nM	P1, 250 nM
#2 primers with deoxy- INOSINE	F2A, 500 nM	R2A, 250 nM +R3A, 250 nM +R4A, 250 nM	P1, 250 nM
#3 primers without deoxy-INOSINE	F2B, 500 nM	R2B, 250 nM +R3B, 250 nM +R4B, 250 nM	P1, 250 nM
#4 primers from Grywna (2010)	F1, 500 nM	R1, 500 nM	P2, 250 nM
#5 primers with deoxy- INOSINE	F2A, 500 nM	R2A, 250 nM +R3A, 250 nM +R4A, 250 nM	P2, 250 nM
#6 primers without deoxy-INOSINE	F2B, 500 nM	R2B, 250 nM +R3B, 250 nM +R4B, 250 nM	P2, 250 nM

Each mix contains the forward primer, up to 3 reverse primers and a probe as mentioned. Combination of 3 reverse primers accounts for nucleotide variation by limiting the use of degenerated primers