Table 2 Metabarcoding assay information for subterranean eDNA surveys on Christmas Island.

From: Under the karst: detecting hidden subterranean assemblages using eDNA metabarcoding in the caves of Christmas Island, Australia

PCR assay

Target taxa

Primer name

Oligonucleotide sequence (5′–3′)

Target length (bp)

Annealing temp (°C)

Primer reference

16S Fish (short)

Bony Fish

16S_FishSyn_Short forward

GACGAGAAGACCCTGTGGAGC

70–140

55

Nester et al.27

16S_FishSyn_Short reverse

CCGYGGTCGCCCCAAC

16S Crustacean

Crustacean

Crust16S_F (short)

GGGACGATAAGACCCTATA

90–213

51

Berry et al.28

Crust16S_R (short)

ATTACGCTGTTATCCCTAAAG

18S Universal

Eukaryotes

18S_1F

GCCAGTAGTCATATGCTTGTCT

336–423

52

Pochon et al.29

18S_400R

GCCTGCTGCCTTCCTT

  1. Three PCR primer sets: 16S Fish (short), 16S Crustacean and 18S Universal corresponding to the mitochondrial 16S rRNA and nuclear 18S regions, were applied to all collected water and sediment samples. In the primer name, “F” refers to the forward primer and “R” to the reverse primer.
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