A 611 base pair (bp) fragment of the mitochondrial cytochrome oxidase I (mtCOI
) gene was sequenced from 44 adult copepods.
PCR amplifications for the mtCOI
gene were also performed using the Btab-Uni primer set described by Shatters et al.
Upon obtaining 660 bp of unambiguous mtCOI
sequence for one exotic Philippine mussel, Basic Local Alignment Search Tool searches to GenBank indicated that a portion of this initial sequence was 100% identical to bases 1-619 of Mytella charruana mtCOI
haplotype A (accession no.
A portion of mtCOI
was amplified with a polymerase chain reactionusing primers, BaQ F (5/GAAGCAACGACTACTTACAA-3/), BaQ R (5/TTCTCGGCGTTTTTACCAA-3/) and BaB F (5/ CCACTATAATTATTGCTGTTCCCACA-3/),L2N 3014R(5/TCCAATGCACTAATCTGCCCATCTGCCCATATTA-3/).The PCR was performed using 5pl of DNA extract in 25pl of reaction mixture containing 4.5 [mu]l of distilled water, 12.5 [mu]l of Taqmix (buffer, Mg[cl.sub.2], dNTPs, TaqDNA polymerase)(MBI Fermentas, Life Sciences, York, UK), 1.5 [mu]l each of forward and reverse primer and 5[mu]l of DNA.
Most of the sequence data are from a single mitochondrial locus, sequenced at two distinct regions (mtCOI
Siguiendo la metodologia para el analisis y comparacion de la secuencia parcial del gen mtCOI
Polymerase chain reaction (PCR) was performed with mtCOI
primers F-C1-J-2195 (5'-TTGATTTTTTGGTCATCCAGAAGT-3') and R-L2N-3014 (5'-TCCAATGCACTAATCTGCCATATTA-3') (Simon et al.
Vestimentiferan-specific primers COIF and COIR (Nelson and Fisher, 2000) were used to amplify a 1200-base pair (bp) fragment of mtCOI
. However, to be consistent with previous studies of mtCOI
diversity in Osedax and other deep-sea polychaetes (Rouse et al., 2004), all estimates obtained for this study were based on a 540-bp region from the 5'-end of this gene.
Among the genetic markers mtCOI
has gained widespread prominence during the past eight years (Dai et al.
However, these species did not form a clade on the phylogenetic trees constructed based on nuclear 28S rRNA or the mitochondrial cytochrome oxidase I (mtCOI
) and 16S rRNA genes.
Although other molecular data sets are available (e.g., Sotka et al., 2004), estimates of the substitution rate [mu] have only been calculated for this portion of the mtCOI
locus in balanoid barnacles (see Materials and Methods).
spiniferus using molecular marker (mtCOI
gene), morphological and acoustic analysis to distinguish it from A.