Table 3.
Target organism | Oligo | Primer/probe | Target loci | Sequence 5′–3′ | Amplicon length | Annealing temperature |
---|---|---|---|---|---|---|
Chattonella verruculosa | 227 | 58° C | ||||
ChattaqFor | Forward | 18S rRNA | CCGTAGTAATTCTAGAGCTAATACRTGCA | |||
ChattaqRev | Reverse | 18S rRNA | AATTCTCCGTTACCCGTTAAAGCCAT | |||
CVerrProbe | Probe | 18S rRNA | [6~FAM]AATGGCGCGCAAGCG-TGTATTATG[TAMRA~Q] | |||
C. marina | 224 | 60° C | ||||
C. antiqua | CmarinaFor | Forward | ITS2 rRNA | GGTAGTTGCCGTACATTTTGCTCTT | ||
C. ovata | CmarinaRev | Reverse | ITS2 rRNA | AAAAGTGGATTCAGCCGAAGCTTC | ||
CmarinaProbe | Probe | ITS2 rRNA | [6~FAM]TTGAGTTCAACGGGCGTG-GTAGC[BHQ~6~FAM] | |||
C. subsalsa | 156 | 60° C | ||||
SubsalsaFor | Forward | ITS2 rRNA | TTGGATTCCGACGGGC | |||
SubsalsaRev | Reverse | ITS2 rRNA | ATATGCTTAAATTCAGCGGGTTTT | |||
SubsalsaProbe | Probe | ITS2 rRNA | [6~FAM]TTCGGCCAAGCACACAT-CCTC[TAMRA~6~FAM] | |||
Fibrocapsa japonica | 167 | 57° C | ||||
Fjap490For | Forward | 28S rRNA | TGAAAACGGCCCGTACACA | |||
Fjap657Rev | Reverse | 28S rRNA | CGGGAACAGCTCATGATGT | |||
Fjap578Probe | Probe | 28S rRNA | [6~FAM]CGGCTGGACACGCTTCTG[BHQ1~Q] | |||
Heterosigma akashiwo | 228 | 62° C | ||||
Haka127For | Forward | 28S rRNA | AAAGGTGCGTGCTCAGTCGTGGT | |||
Haka355Rev | Reverse | 28S rRNA | CAAAAGTCTTTTCATCTTTCCCT | |||
Haka222Probe | Probe | 28S rRNA | [5TET]TACGAGCCGTTTCCGACGA[BHQ1~Q] |
Loci were chosen based on ability to distinguish target species from others. Amplicon length and annealing temperatures are provided.