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. 2013 Oct 9;8(10):e75606. doi: 10.1371/journal.pone.0075606

Figure 4. Blind PCR-RFLP assay with eighteen pearls of unknown identity.

Figure 4

(A) PCR products of 575 bp (Pinctada margaritifera), 571 bp (P. maxima) and 590–91 bp (P. radiata) obtained with ITS2 universal primers (5.8S-F and 28S-R) and (B) of 335 bp obtained with 28S-R and the P. margaritifera specific primer ITS2-Marg-F. (C) RFLP patterns of ITS2 gene fragments (from A) obtained after digestion with RsaI. MW: molecular weight size marker, 100 bp DNA ladder; lanes 1–18: pearl isolates; lanes 19–20: DNA extraction negative controls; lane 21: PCR negative control; lanes 22–23: P. radiata and P. margaritifera positive controls; lanes 24–26: P. radiata, P. margaritifera and P. maxima positive controls showing ITS2 PCR products (upper gel) and ITS2-RFLP patterns (lower gel).