PacBio of Bunonema sp. JU3390: mixed stage and sex population
Promega Wizard HMW DNA Extraction Kit (cat. no. A2920) was used for DNA long-read sequencing using the manufacturer's protocol with minor modifications. Worm cuticles were disrupted through routine freeze/thaw cycles, alternating between -80C for five minutes and 37C until thawed, with brief vortexing between cycles, repeated five times. All centrifugation steps were performed at 4C, and alcohols were kept on ice until required. An additional incubation of 25 minutes at 65C was incorporated at the lysis step. Samples were subsequently given to the University of Miamis John P. Hussman Institute for Human Genomics Sequencing Core Facility (RRID:SCR_017828) for PacBio sequencing