Isolation and characterization of 16 highly polymorphic tetranucleotide microsatellite DNA markers in Paa spinosa

The giant spiny frog, Paa spinosa is a vulnerable species. By using a modified FIASCO (Fast Isolation by AFLP Sequences Containing repeats) method, we isolated and characterized 16 polymorphic tetranucleotide microsatellite loci for P. spinosa. These loci had high polymorphism when screened in 33 randomly caught individuals. The average number of alleles for 16 loci was 7.13 (range 2-14). Observed and expected heterozygosities were 0.152-0.818 and 0.144-0.918, respectively, and did not deviate from Hardy-Weinberg expectations. There was no evidence for scoring error due to stutter, null alleles, or large allele dropout. No significant association between pairs of loci was found after Bonferroni correction. These polymorphic loci will be effective for evaluating genetic diversity, population structure, gene flow and determining paternity in this frog, and also for conservation management.