Single-step species identification of bivalve larvae using multiplex polymerase chain reaction |
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Authors: | M P Hare S R Palumbi C A Butman |
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Institution: | (1) University of Maryland, Dept. of Biology, College Park, MD, USA 20742-4415 Fax: 001 301 314 9358 e-mail: mh285@umail.umd.edu, US;(2) Applied Ocean Physics & Engineering Department, Woods Hole Oceanographic Institution, Mail Stop 12, Woods Hole, Massachusetts 02543, USA, US |
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Abstract: | One of the biggest obstacles to studying recruitment variation in marine bivalves is the need to collect and process large
numbers of plankton samples. Larval bivalves are notoriously difficult, if not impossible, to identify to species using morphological
criteria alone. Remote time-series collections could satisfy the sampling challenge, but efficient identification techniques
must be developed to obtain species-specific data. Thus, we have developed a multiplex polymerase chain reaction (PCR) identification
assay in which a single reaction is capable of accurate and efficient discrimination of five target bivalve species based
on the size of cytochrome oxidase I products. The assay was tested with cultured and field-sampled larvae as well as adult
genomic DNAs. Using a single whole larva as template, multiplex PCR reactions were capable of discriminating among the commercially
important bivalves: Mercenaria mercenaria, Argopecten irradians, Mulinia lateralis, Spisula solidissima and Mya arenaria. Overall accuracy was 92%, including very few false positives. The efficiency of this assay stems from its ability to discriminate
multiple target species with a single molecular step that ultimately can be automated to process large numbers of larvae.
Received: 30 March 2000 / Accepted: 10 July 2000 |
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