PacBio CCS sequencing of E. vannus: isolated germline genomic DNA
Total genomic DNA was extracted using phenol-chloroform and digested using 40 units of the methylation-sensitive DpnI restriction endonuclease (Cat. R0176, NEB) for 3 hours to deplete bacterial DNA. Germline DNA molecules longer than 10 Kb were isolated from the processed DNA using a BluePippin system. The HiFi SMRTbell sequencing library was constructed using SMRTbell Express Template Prep Kit (2.0) with ~15 Kb segments. The isolated DNA was sequenced with PacBio long-read system in circular consensus sequencing (CCS) mode.