Table 2 Design of 13 candidate qPCR assays in ascending order by template copy number
qPCR assay | Fluorescence | Forward primer (FP), reverse Primer (RP) and Probe sequences (5′ → 3′) |
|---|---|---|
crt | MGB TaqMan | FP: GACACCGAAGCTTTAATTTAC RP: GCAGAAGAACATATTAATAGGAA Probe: 5′-HEX/TTAGATGCCTGTTC/3′-MGBEc/ |
crt76K crt76T | MGB TaqMan | FP: TGGTAAATGTGCTCATGTGTTT RP: AGTTTCGGATGTTACAAAACTATAGT Probe(K76): 5′-HEX/TGTGTAATGAATAAAATTTTTGCTAA/3′-MGBEc/ Probe(T76): 5′-6-FAM/TGTGTAATGAATACAATTTTTGCTAA/3′-MGBEc/ |
ldh(a) | Standard TaqMan | FP: ACGATTTGGCTGGAGCAGAT RP: TCTCTATTCCATTCTTTGTCACTCTTTC Probe: 5′-HEX/AGTAATAGTAACAGCTGGATTTACCAAGGCCCCA/3′-IABkFQ/ |
ldh(b) | MGB TaqMan | FP: TGGTCATATTAAGAAGAATTGTC RP: CTGAGATATGTAATACTTCAATC Probe: 5′-HEX/CCATAACATCTACT/3′-MGBEc/ |
18SrRNA(a) | Standard TaqMan | FP: CTTTTGAGAGGTTTTGTTACTTTGAGTAA RP: TATTCCATGCTGTAGTATTCAAACACAA Probe: 5′-HEX/TGTTCATAACAGACGGGTAGTCATGATTGAGTTCA/3′-IABkFQ/ |
18SrRNA(b) | MGB TaqMan | FP: ATTGCTTTTGAGAGGTTTTGTTACTTT RP: GCTGTAGTATTCAAACACAATGAACTCAA Probe: 5′-HEX/CATAACAGACGGGTAGTCAT/3′-MGBEc/ |
r364(a) | PET | FP: ACCCCTCGCCTGGTGTTTTT RP/Probe: 5′-HEX/aggcgcatagcgcctggTCGGGCCCCAAAAATAGGAA |
cytB | LNA TaqMan | FP: TACTAACTTGTTATCCTCTATTCCAGTAGC RP: CCTTTAACATCAAGACTTAATAGATTTGGA Probe: 5′-HEX/+ GTG + CTA + CCA + TGT + AAA + TGTAA/3′-IABkFQ/ |
coxI | MGB TaqMan | FP: GTCACGCAATATCAATATACTG RP: CGATCTCTTGTATGGTAATAGG Probe: 5′-HEX/ATAGAACTCCAGGC/3′-MGBEc/ |
varATS | MGB TaqMan | FP: CCCATACACAACCAAYTGGA RP: TTCGCACATATCTCTATGTCTATCT Probe: 5′-HEX/TRTTCCATAAATGGT/3′-MGBEc/ |
r364(b) | MGB TaqMan | FP: AGTCCATTTTCCCCTAGC RP: GACCATATAGTAAGTGACCCA Probe: 5′-HEX/AATTGACATGCACT/3′-MGBEc/ |
TARE-2 | dsDNA-dye | FP: CTATGTTGCACTTACATGCAYAAT RP: TGACCTAAGAAGTAVAATAATGATGA |