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MgC12; 1.5 mM dNTPs (AB); 0.5 µM each, forward and reverse primers. Forward <br />primers were labeled with one of four fluorescent dyes (6-FAM, PET, NED, VIC; AB). <br />We added 2 µL of genomic DNA to each reaction; the concentration of DNA varied. The <br />reactions were amplified in GeneAmp® PCR System 9700 thermal cyclers (AB) using <br />the following thermal profile: 95 °C for 9 min, followed by 33 cycles of 94°C for 1 min, <br />56°C for 45 sec (decreasing by -0.2°C per cycle) and. 72°C for 1 min, with a final <br />extension of 72°C for 45 min. The PCR products were visualized on an AB 3130x1 <br />genetic analyzer. GeneScanTm 500 LIZTm size standard (AB) was used. Composite <br />genotypes for individual fish were compiled with GeneMapper 3.5 software (AB). <br />Approximately 10% of the DNA samples on each plate were duplicated. and scored <br />separately to ensure quality control. <br />Statistical analyses <br />Genepop on the Web (version 3.4) (updated from Raymond and Rousset 1995) <br />was used to test for genotypic linkage disequilibrium and numbers of observed and <br />expected heterozygotes by sample and. locus. Each pair of loci in each sample was <br />checked for linkage disequilibriumm with Fishers' exact test; Markov chain parameters <br />were 1000 dememorization steps, 100 batches, and 10,000 iterations per batch. Statistical <br />significance was set at a = 0.05 and adjusted for multiple tests using the standard <br />Bonferroni method (Rice 1989). <br />FSTAT (version 2.91, Goudet 1995) was used to calculate allele frequencies per <br />locus and allelic richness per locus and sample, Fis per locus and sample, global values of <br />F-statistics (Wright 1951) and. the variance components from which they were estimated..