Discovery and genetic characterization of single cohort adult colonies with male aggregations, and preliminary evidence for lekking in a Malagasy kite spider (Isoxya, Gasteracanthinae)

Genomic DNA was isolated from 80 individuals representing nine web colonies using the Qiagen DNeasy Blood and Tissue kit (Qiagen). SNP data was generated using the 3RAD technique detailed in Bayona-Vásquez et al. (2019). We used enzymes EcoRI, MspI, and ClaI following Petersen et al. (2012) and Burns et al. (2017), and adapters provided by the University of Georgia (BadDNA@UGA). Paired-end 150 base pair reads were generated for the 3RAD library with Nextseq at the University of California, Davis DNA Technologies Core. Further detail regarding our 3RAD procedure is given in Newton et al. (2020). Read demultiplexing, filtering, and clustering were performed with ipyrad (Eaton and Overcast, 2020). For each end of each locus, maxima of six uncalled bases, ten heterozygous sites, 25 SNPs, and ten indels were allowed. The clustering threshold was set to 0.9, and locus occupancy was set to a minimum of 50%.