As a first step to establish the genetic structure of the sea urchin Echinometra lucunter lucunter throughout the Caribbean Sea, 26 microsatellite loci were isolated using Illumina paired-end sequencing, Next Generation Sequencing (NGS). We successfully optimized 17 loci for genotyping and the variation tested for 23 individuals from the Caribbean Sea and Tropical Eastern Atlantic Ocean. The allele number per locus (Na) ranged from four to 24, the observed heterozygosity (Ho) from 0.682 to 1, and the expected heterozygosity (He) from 0.609 to 0.9304. We detected no linkage disequilibrium between pairs of loci. These microsatellites will be used for the first time to detect the influence of marine barriers to genetic flow in the sea urchin E. lucunter lucunter throughout the Caribbean Sea. These new validated markers will be essential for current conservation and connectivity studies across the Caribbean Sea and the Atlantic Ocean.