Table 3 Primers and probes used for real time RT-PCR in this study
From: A one step real-time RT-PCR assay for the quantitation of Wheat yellow mosaic virus(WYMV)
Target | Accession number | Primer names | Sequence | Tm(°C) | GC (%) | Nucleotide position |
|---|---|---|---|---|---|---|
CP full-length | AJ130983 | WCP-F: | 5′GCAGCTGACACACAAAC 3′ | 41 | 53 | 1156–1172 |
WCP-R: | 5′GGTTAGTTCTGGGTGTCC 3′ | 43 | 56 | 2017–2034 | ||
CP probe | AJ130983 | WF: | [FAM]-GCCGCGTACGCTTTTGACT-[TAMRA] | 55 | 58 | 1843–1861 |
WT: | [FAM]-CTTTGTCCCCCGCCCGTGG-[TAMRA] | 64 | 74 | 1863–1881 | ||
WR: | [FAM]-TTCGATGTCCTGTGGGTTCA-[TAMRA] | 53 | 50 | 1883–1902 | ||
Wheat β-actin | AB181991 | AF: | [FAM]-TCCAATCTATGAGGGATACACGC-[TAMRA] | 55 | 48 | 263–286 |
AT: | [FAM]-TTCCTCAAGCTATCCTTCGTTTGGACCTG-[TAMRA] | 65 | 48 | 286–314 | ||
AR: | [FAM]-TCTTCATTAGATTATCCGTGAGGTC-[TAMRA] | 53 | 40 | 324–348 | ||
Wheat 18S rRNA | BJ217595 | SF: | [FAM]-CAGGCGATGGGAAA-[TAMRA] | 42 | 57 | 199–212 |
ST: | [FAM]-TCGCTTTGAGTTTGGGCAATTGTGG-[TAMRA] | 65 | 48 | 136–160 | ||
SR: | [FAM]-CATTGGAGGGCAAGTCT-[TAMRA] | 45 | 53 | 283–299 |