Shotgun metagenomic data were generated for 138 samples (74 vaginal and 64 penile) as described previously (84 (link)). Swabs were resuspended in 1 mL of Amies transport medium. Genomic DNA was extracted using 200 µL of resuspended material and the MagAttract PowerMicrobiome DNA/RNA Kit (Qiagen) with bead-beating using TissueLyser II. Sequence libraries were generated from extracted DNA using the Illumina Nextera XT Flex kit. Procedures were performed according to manufacturer instructions and automated on the Hamilton STAR robotic platform. Libraries were sequenced on an Illumina NovaSeq 6000 (150 bp, paired-end) at Maryland Genomics at UMSOM. Host reads were identified and removed using BMtagger and the GRCh38 human genome as reference (91 ). Non-host reads were processed using fastp (v.0.21) (92 (link)). Metagenome taxonomic compositions were determined via mapping to VIRGO (59 (link)), and CSTs were assigned using VALENCIA (2 (link)). VALENCIA is a supervised classifier for the vaginal microbiome. The penile CST assignments we observed are consistent with prior unsupervised clustering results (25 (link), 27 (link)– (link)29 (link)), so we feel it is reasonable to use VALENCIA with penile samples in STING.