Long-read Sequencing of Genomic DNA

DNA was extracted from overnight cultures (OD_600nm = 1.5) using PowerSoil DNA Isolation Kit (Qiagen, Hilden, Germany). After quantification by gel electrophoresis and fluorimetric assay (Qubit, Thermo Fisher Scientific, Waltham, MA, USA), we followed the procedure already reported in [25 (link)] for fragmenting DNA with g-TUBE (Covaris Inc., Woburn, MA, USA) to an average 15 kbp size and preparing the sequence library using the Pacific Biosciences SMRTbell Express Template Prep Kit 2.0 (Pacific Biosciences, Menlo Park, CA, USA). Sequencing was performed on a Sequel apparatus (Pacific Biosciences, Menlo Park, CA, USA) by SMRT technology [21 (link)], using Sequel Sequencing Kit 3.0.