Loci | Primer sequences | PCR conditions |
---|---|---|
18S rRNA gene (region 1) | M13F-18S ai (5′-TGTAAAACGACGGCCAGT CCTGAGAAACGGCTACCACATC-3′) M13R-18S bi (5′-CAGGAAACAGCTATGAC GAGTCTCGTTCGTTATCGGA-3′) [55] | Denaturation at 94 °C for 30 s, annealing at 52 °C for 45 s and extension at 68 °C for 1 min 15 s, for 34 cycles |
18S rRNA gene (region 2) | M13F-NF1 (5′-TGTAAAACGACGGCCAGT GGTGGTGCATGGCCGTTCTTAGTT-3′) M13R-18Sr2b (5′-CAGGAAACAGCTATGAC TACAAAGGGCAGGGACGTAAT-3′) [56] | Denaturation at 94 °C for 30 s, annealing at 54 °C for 45 s and extension at 68 °C for 30 s, for 34 cycles |
COI | M13F-CIJ1718 (5′-TGTAAAACGACGGCCAGT GGAGGATTTGGAAATTGATTAGT-3′) M13R-CIN2191 (5′-CAGGAAACAGCTATGAC CAGGTAAAATTAAAATATAAACTTCTGG-3′) [43] | Denaturation at 94 °C for 30 s, annealing at 46 °C for 45 s and extension at 68 °C for 40 s, for 34 cycles |
28S rRNA gene (D1D2) | M13F-C′1 (5′-TGTAAAACGACGGCCAGT ACCCGCTGAATTTAAGCAT-3′) M13R-D′2 (5′-CAGGAAACAGCTATGAC TCCGTGTTTCAAGACGGG-3′) [34] | Denaturation at 94 °C for 30 s, annealing at 50 °C for 45 s and extension at 68 °C for 50 s, for 34 cycles |