TABLE 2.

Primers and standard sequences used for one-step RT-qPCR to quantify viruses bound to PGM-MBs

Target gene	Primer name	Sequence (5′–3′)	Position in the gene (nt)	Amplicon size (bp)	Reference	RT-qPCR conditions
RV strain OSU NSP3a	JVKF	CAGTGGTTGATGCTGAAGAT	17–37	122	72	1 cycle at 50°C for 10 min and 95°C for 1 min; then 40 cycles at 95°C for 10 s and 60°C for 30 s
	JVKR	TCATTGTAATCATATTGAATACCCA	122–138
TV NSPb	TV-NSP-qPCR-F2	GGCAGCTGGGAAGAAATCTG	2774–2793	111	15	1 cycle at 50°C for 10 min and 95°C for 1 min; then 40 cycles at 95°C for 10 s and 60°C for 30 s
	TV-NSP-qPCR-R2	CCTGCTGTGTGAATGCCTAC	2865–2884

^aqPCR standard sequence (5′–3′) acquired from GenBank accession number DQ146697.1: GGCTTTTAATGCTTTTCAGTGGTTGATGCTCAAGATGGAGTCTACTCAACAGATGGCATCTTCTATTATTAACTCTTCTTTTGAAGCTGCAGTTGTCGCTGCAACTTCTACATTAGAATTAATGGGTATTCAATATGATTATAATGAAGTATACACTAGAGTTAAAAGTAAGTTTGATTTTGTAATGGATGATTCTGGTG. The forward (F) primer attaches to the sequence indicated in boldface, and the reverse primer (R) attaches to the complementary strand indicated by underlining.

^bqPCR standard sequence (5′–3′) acquired from GenBank accession number EU391643.1: CCGTGGTTGTGCGCAGTATTGGAAACACAAACATTGCTGGGAAATTCCTCAACGTCTTCACAGGTACAGTTGTGGCAGCTGGGAAGAAATCTGACGGCCTCGGGTCTGAACCAGGAGACTGTGGCTCACCATATCTTAAATTTGTTAATGGAAAACCAACTCTTGTAGGCATTCACACAGCAGGCAGCTACACTACCAACCAGGTTGCAGGCTTAGTGATACCTTCTAGATTCAACCTTG. The forward (F) primer attaches to the sequence indicated in boldface, and the reverse primer (R) attaches to the complementary strand indicated by underlining.