16S amplicon sequencing from control or PANDAS subjects
The V4 hypervariable region of the 16S rRNA gene was PCR-amplified using the 515-F and 806-R primer pair with Illumina adapters and a unique 12 base pair (bp) barcode for each sample. PCR product was quantified with a Quant-iT PicoGreen dsDNA Assay Kit (Invitrogen). DNA products were then pooled in equimolar concentration; products longer than 200 bp were selected using a QIAseq beads and sequenced on a 2x150 bp Illumina MiSeq platform (Illumina)