title: Neogobius melanostomus microsatellite sequences: tissues and eDNA samples
design: To investigate genetic diversity in eDNA and tissue samples, we amplified a panel of 28 microsatellite markers for Round Gobies. Primer pairs were grouped into multiplexes and eDNA samples were amplified with each multiplex assay in triplicate to account for PCR stochasticity. Equal volumes (5 L) of each technical replicate and multiplex were combined for each sample and uniquely indexed in a second-stage PCR using Illumina Nextera XT tags. We pooled the final amplicons of each sample and negative control (field, extraction, and PCR) in equal volumes and purified the eDNA library with Agencourt AMPure XP beads. We sequenced libraries at Cornell University's Institute of Biotechnology Genomics Facility on the Illumina MiSeq platform with the MiSeq v2 500 bp kit (paired-end 2  250 bp).