Mp. Hare et al., Single-step species identification of bivalve larvae using multiplex polymerase chain reaction, MARINE BIOL, 137(5-6), 2000, pp. 953-961
One of the biggest obstacles to studying recruitment variation in marine bi
valves 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 collecti
ons could satisfy the sampling challenge, but efficient identification tech
niques must be developed to obtain species-specific data. Thus, we have dev
eloped 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 p
roducts. The assay was tested with cultured and field-sampled larvae as wel
l as adult genomic DNAs. Using a single whole larva as template, multiplex
PCR reactions were capable of discriminating among the commercially importa
nt 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 abili
ty to discriminate multiple target species with a single molecular step tha
t ultimately can be automated to process large numbers of larvae.