Isolation and characterisation of 12 polymorphic microsatellite loci for the threatened mound-building malleefowl, Leipoa ocellata (Aves:Megapodiidae)

Abstract

Using 454 pyrosequencing and genomic enrichment techniques we developed 12 polymorphic markers for the endangered megapode, the malleefowl (Leipoa ocellata). Pyrosequencing on a 454 instrument resulted in 65536 reads, with 3469 containing microsatellite repeats. Of these, 232 contained unique flanking sequences and had more than 8 repeat motifs. We chose 13 loci based on reliability of amplification and, from these, 12 unlinked loci were selected for genotyping. In a single population (n≤19), the 12 markers were moderately polymorphic (number of alleles per locus range≤3-7) and showed moderate to high levels of heterozygosity (0.285-0.882). Nine microsatellite primer pairs developed from the..