Genome Sequencing of S. dysgalactiae subsp. equisimilis

The genomes of the S. dysgalactiae subsp. equisimilis strains were sequenced with methods described previously for S. pyogenes (17 (link), 38 (link), 54 (link), 61 (link), 62 (link)). Briefly, strains were grown at 37°C with 5% CO₂ on tryptic soy agar with 5% sheep blood (Becton, Dickinson, Franklin Lakes, NJ) or in Todd-Hewitt broth with 2% (wt/vol) yeast extract (THY; Difco Laboratories, Franklin Lakes, NJ). Chromosomal DNA for Illumina short-read sequencing was isolated with the RNAdvance viral kit (Beckman Coulter, Brea, CA) and a BioMek i7 instrument (Beckman Coulter). Libraries were prepared with the NexteraXT kit (Illumina, San Diego, CA) and sequenced with a NovaSeq instrument (Illumina) using a 2 × 250-bp protocol. Chromosomal DNA for long-read sequencing was isolated with a DNeasy blood and tissue kit (Qiagen, Germantown, MD). Libraries were prepared with either a native barcoding kit or rapid barcoding kit (Oxford Nanopore Technologies, United Kingdom) and sequenced with a GridION instrument using version R10.4 or R9.4.1 flow cells (Oxford Nanopore Technologies), respectively.