Genome Sequencing of A. arguta cv. 'Longcheng No.2'

Green wood cuttings of A. arguta cv. ‘Longcheng No.2’ were picked and grown in a tissue culture incubator at Anhui Agricultural University, Anhui Province, China, under 25 °C, 12-/12-h days. Fresh young healthy leaves were collected from 3-week-old branches, quickly frozen with liquid nitrogen and then stored at –80 °C for PacBio HiFi and Hi-C sequencing. High molecular weight genomic DNA (gDNA) was extracted separately from each leaf tissue sample using a slightly modified cetyltrimethylammonium bromide (CTAB) method (Allen et al. 2006 (link)). The quality and quantity of the isolated gDNAs were evaluated with an Agilent 2100 Bioanalyzer (Agilent Technologies, CA, USA) and a Qubit fluorometer instrument (Thermo Fisher Scientific, MA, USA), respectively. For PacBio HiFi sequencing, a standard SMRTbell library was prepared with 50 μg gDNA using the SMRTbell Express Template Prep Kit 2.0 according to the manufacturer's instructions. SMRTbell libraries were then sequenced on the PacBio Sequel II system (Pacific Biosciences, CA, USA). The Hi-C sequencing library was prepared and sequenced based on a previously published protocol (Rao et al. 2014 (link)).