title: WGS of Bursaphelenchus mucronatus: mix stage
design: Total genomic DNA was extracted from nearly 20,000 isolates of mixed age using MagAttractHMW DNA Kit (QIAGEN, Valencia, CA, USA) according to the manufacturer's protocol. Agarose gel electrophoresis was used to check the integrity of genomic DNA molecules and the main band was around 20kb, which satisfied the requirement for PacBio long read library construction. SMRTbell Template Prep Kit was subsequent used to construct the library for the PacBio RS II platform (Pacific Biosciences of California, Menlo Park, CA, USA). A total of 24 single-molecule real-time cells were produced with P6/C4 chemistry, generating 720 million reads with a total length of 24-Gb subreads with a coverage of 110 of the B. mucronatus genome. Meanwhile, KAPA Hyper Prep Kit (KK8504) was used to construct the paired-end library for HiSeq 4000 platform (Illumina, San Diego, CA, USA) with insert sizes of 300 bp, which was used for genome survey sequencing.