Cyrtotrachelus buqueti Genome sequencing and assembly. Cyrtotrachelus buqueti

Fresh Mixed tissue samples were obtained from a male individual of C. buqueti using a Universal Genomic DnNA Kit CW2298 (Cwbiotech Co., Ltd., Beijing, China) according to the manufacturer’s operating manual. Next, 1% agarose gel electrophoresis of the extracted genomic DNA was performed, after which the concentration was quantified using a Qubit 3 fluorometer (Thermo Fisher, Waltham, MA, USA). Before genome sequencing, genomic libraries (SMRTbell libraries) for Pacific Biosciences (PacBio) long-read sequencing were constructed by shearing DNA into ~20 kilobase (kb) fragments using a Covaris g-TUBE (KBiosciences part No. 520079), after which they were enzymatically repaired and nine 20-kb SMRTbell libraries were constructed using a DNA Template Prep Kit 1.0 (PacBio part No. 100-259-100). The DNA fragments size distribution was subsequently estimated using a Bioanalyzer 2100 12K DNA Chip assay (Agilent part No. 5067-1508), after which the templates were size-selected using a BluePippin (Sage Science, Inc., Beverly, MA, USA) to enrich large DNA fragments (> 10 kb). An Agilent Bioanalyzer 12 kb DNA Chip (Agilent Technologies, Santa Clara, CA, USA) and Qubit fluorimeter (Invitrogen, Carlsbad, CA, USA) were employed to inspect the quality of the fragments and quantify the library. Binding Kit 2.0 (PacBio part No. 100-862-200) was used to construct a SMRT BellPolymerase Complex as described in the protocols. Genome sequencing was conducted at Biomarker Technologies Corporation (Beijing, China) using a PacBio Sequel sequencer (Pacific Biosciences, Menlo Park, CA, USA). Samples were loaded and sequenced on PacBio SMRT cells v3.0 (PacBio part No. 100-171-800) of the Sequel instrument, which obtained 360 min per SMRT cell in one movie. Finally, MagBead loading (PacBio part No. 100-125-900) was introduced to improve enrichment of the large fragments.