PacBio Hifi of Micrasema longulum
Tissue was used for high molecular weight DNA extraction using an adaptation of the protocol by Miller et al. (1988). Final DNA purity and concentrations were measured using NanoPhotometer (Implen GmbH, Munich, Germany) and Qubit Fluorometer (Thermo Fisher Scientific, Waltham, MA).  Two samples were pooled to construct one multiplexed SMRTbell library following the instructions of the SMRTbell Express Prep kit v2.0 with Low DNA Input Protocol (Pacific Biosciences, Menlo Park, CA). The input DNA for each sample was 77 ng (Micrasema setiferum) and 489 ng (Micrasema longulum), respectively. The final library was loaded at an on-plate concentration of 80 pM using diffusion loading. One SMRT cell sequencing run was performed on the Sequel System II  in CCS mode using 30 hour movie time with 2 hours pre-extension and sequencing chemistry V2.0.