476 
Microsatellite primers for red drum 
(Sciaenops ocellatus ) 
Email address for J.R. Gold: goldfish@tamu.edu 
1 Center for Biosystematics and Biodiversity 
Texas A&M University, TAMU-2258 
College Station, Texas 77843-2258 
2 USDA/ARS National Center for Cold and Cool Water Aquaculture 
11861 Leetown Road 
Kearneysville, West Virginia 25430 
Sten Karlsson 1 
Mark A. Renshaw 1 
Caird E. Rexroad III 2 
John R. Gold (contact author ) 1 
In this note, we document polymerase- 
chain-reaction (PCR) primer pairs 
for 101 nuclear-encoded microsatel- 
lites designed and developed from a 
genomic library for red drum ( Sciae - 
nops ocellatus ). Details of the genomic 
library construction, the sequencing 
of positive clones, primer design, and 
PCR protocols may be found in Karls- 
son et al. (2008). The 101 microsatel- 
lites (Genbank Accession Numbers 
EU015882-EU015982) were amplified 
successfully and used to genotype 24 
red drum obtained from Galveston 
Bay, Texas (Table 1). A total of 69 of 
the microsatellites had an uninter- 
rupted (perfect) dinucleotide motif, 
and 30 had an imperfect dinucleo- 
tide motif; one microsatellite had an 
imperfect tetranucleotide motif, and 
one had an imperfect and compound 
motif (Table 1 ). Sizes of the cloned 
alleles ranged from 84 to 252 base 
pairs. A ‘blast’ search of the Genbank 
database indicated that all of the 
primers and the cloned alleles were 
unique (i.e., not duplicated). 
Summary genotypic data, based on 
22-24 assayed red drum, also are 
given in Table 1 and include number 
(and size range) of alleles detected, 
observed and expected heterozygos- 
ity, and probability values from tests 
for conformity to Hardy-Weinberg 
equilibrium expectations. One mic- 
rosatellite (Soc734) was monomor- 
phic; the number of alleles detected 
at the remaining 100 (polymorphic) 
microsatellites ranged from 2 to 26. 
Estimates of observed and expected 
heterozygosity and tests for confor- 
mity to Hardy-Weinberg and geno- 
typic equilibrium expectations were 
performed with Genepop (Raymond 
and Rousset, 1995). Observed het- 
erozygosity (polymorphic microsat- 
ellites) ranged from 0.042 (Soc706) 
to 1.000 (11 microsatellites, Table 1) 
and averaged (±standard deviation 
[SD] ) 0.775 ±0.211; expected hetero- 
zygosity ranged from 0.042 (Soc706) 
to 0.971 (Soc636) and averaged 0.806 
±0.201. After Bonferroni correction 
(Rice, 1989), genotypes at 99 of the 
polymorphic microsatellites did not 
differ significantly from Hardy-Wein- 
berg equilibrium expectations. At one 
locus, Soc 706, there were only two al- 
leles, one of which was observed only 
in a heterozygote; this microsatellite 
was not tested for Hardy-Weinberg 
equilibrium. Analysis with MICRO- 
CHECKER (Van Oosterhout et al., 
2004) indicated the possible occur- 
rence of null alleles at nine of the 
microsatellites, and single base-pair 
shifts (i.e., alleles differing by only 
a single base pair) were observed at 
five of the microsatellites (Table 1). 
Tests of genotypic disequilibrium 
were nonsignificant after Bonferroni 
correction. Given that red drum pos- 
sess 24 haploid chromosomes (Gold et 
al., 1988), several of the microsatel- 
lites undoubtedly are linked; deter- 
mination of linkage will await formal 
mapping studies. 
Along with PCR primers for red 
drum microsatellites developed pre- 
viously by O’Malley et al. (2003), 
Saillant et al. (2004), and Karlsson 
et al. (2008), the primers developed 
here will be useful in a variety of 
applications (Liu and Cordes, 2004), 
including analysis of stock structure, 
monitoring and assessment of red 
drum stock enhancement, parentage 
analysis as employed in aquaculture, 
and the generation of a genetic map 
for red drum. A table of the 269 PCR 
primers developed for red drum may 
be found at <http://wfsc.tamu.edu/ 
doc> under the file name “PCR prim- 
ers for red drum (Sciaenops ocellatus) 
microsatellites.” 
Acknowledgments 
We thank C. Abbey for technical 
assistance with the Q-bot (Genetix), 
E. Saillant for assistance in the lab- 
oratory and helpful comments on a 
draft of the paper, and R. Vega for 
encouragement and support. Work 
was supported by the Coastal Conser- 
vation Association and Central Power 
and Light (CCA/CPL) Marine Devel- 
opment Center of the Texas Parks 
and Wildlife Department, the Coastal 
Conservation Association — Texas, and 
the Texas Agricultural Experiment 
Station (Project H-6703). This note 
is number 64 in the series “Genetic 
studies in marine fishes” and contri- 
bution no. 155 from the Center for 
Biosystematics and Biodiversity at 
Texas A&M University. 
Manuscript submitted 10 June 2008. 
Manuscript accepted 12 June 2008. 
Fish. Bull. 106:476-482(2008). 
The views and opinions expressed 
or implied in this article are those 
of the author and do not necessarily 
reflect the position of the National 
Marine Fisheries Service, NOAA. 
