Figure 1.
Sequence of primers and alignment of the conserved regions of 12S ribosomal DNA gene sequences of 64 metazoan species belonging to 23 phyla.
Accession numbers of individuals are denoted in parentheses. Lengths of amplified PCR products without primers are indicated. Hierarchy of NCBI taxonomy database is followed in this figure.
Table 1.
Observed mismatches between the primer pair and the targeted region for all complete mitochondrial genome sequences downloaded from the NCBI Organelle Genome Resources database.
Figure 2.
Nucleotide composition of the primer target regions observed in Nematoda (A and B) and Cnidaria (C and D).
Sequences of the primers are indicated on top of each figure. The nucleotides that matched with the primers are shown stacked in each column. Position of each site from 3′ to 5′ is indicated on bottom from left to right.
Figure 3.
Agarose gel images of the compatibility test using PCR.
The newly designed primers were used in PCR reactions for individuals belonging to various groups within six phyla: Sipuncula- 1 Phascolosoma sp.; Echinodermata- 2 Ophiocoma erinaceus (brittlestar); Chordata- 3 Pseudamiops gracilicauda (fish); Annelida (Polychaeta)- 4 Pherecardia striata, 5 unidentified terebellid species; Arthropoda- 6 Xanthias latifrons (brachyuran crab), 7 Pilodius flavus (brachyuran crab), 8 Liomera sp. (brachyuran crab), 9 Carupa sp. (brachyuran crab), 10 unidentified pilumnid species (brachyuran crab), 11 unidentified xanthid species (brachyuran crab), 12 Calcinus gouti (anomuran crab), 13 Synalpheus sp. (caridean shrimp), 14 Periclimenes sp. (caridean shrimp), 15 unidentified caridean shrimp, 16 unidentified amphipod species; Mollusca- 17 Cypraea helvola (gastropod), 18 Cypraea fimbriata (gastropod), 19 Trivia sp. (gastropod), 20 Erato sandwichensis (gastropod), 21 unidentified haminoeid species (gastropod), 22 Berthellina sp. (gastropod), 23 Chlamys sp. (bivalve), 24 Lima sp. (bivalve), 25 unidentified lucinid species (bivalve). nc-negative control. Good amplifications were observed for all individuals except one bivalve (24).