We characterised 14 new polymorphic microsatellite loci for the endangered lizard Liopholis slateri. Initially, 454 shotgun sequencing was used to identify 46 loci, which were trialled for amplification. Subsequently, 14 of these loci were screened for variation in 21 individuals from scat-derived DNA samples collected from Owen Springs Reserve in central Australia. All 14 loci were polymorphic, with observed heterozygosity ranging from 0.19 to 0.86 and the number of alleles per locus ranging from 2 to 10. These loci will be useful in understanding the genetic variation and connectivity within and among extant L. slateri populations.