Smrtbell template prep kit spv3

The genomic DNA of T. mongolicus was extracted from the leaves using a modified CTAB method [34 (link)]. Quality of the isolated DNA was assessed using NanoDrop-2000 (Thermo Fisher Scientific, Wilmington, DE, USA) and Qubit 3.0 fluorometer (Life Technologies). The genomic DNA was sheared to an average size of 500 bp, and a library with an insert size of 500 bp was prepared using the PairedEnd DNA Sample Prep kit (Illumina Inc., San Diego, CA, USA). The library was sequenced using the Illumina NovaSeq6000 platform (Illumina Inc., San Diego, CA, USA) for genome survey and assessment. For genome sequencing, DNA fragments > 20 kb were selected for library preparation using BluePippin (SAGE). The PacBio library was prepared using the SMRTbell Template Prep Kit-SPv3 following the manufacturer’s instructions (Pacific Biosciences, Menlo Park, CA, USA) and was sequenced on the Pacific Biosciences Sequel system (Pacific Bioscience, CA, USA). The Hi-C library [35 ] was constructed using the HindIII restriction enzyme as per the manufacturer’s instructions (BioMarker Technologies Company) [36 (link)] and was sequenced on Illumina NovaSeq6000 platform for chromosome construction. For RNA-seq used in genome assembly and annotation, libraries were constructed for the root, stem, leaf, and flower of T. mongolicus using a paired-end model and were sequenced on Illumina NovaSeq6000 platform.

Genomic DNA was extracted from N. haitanesis conchocelis using the cetyltrimethylammonium bromide (CTAB) method (Rogers and Bendich 1989 ) and DNA fragments greater than 15 kb were selected using BluePippin (Sage Science, Beverly, MA, USA). The PacBio library was prepared using the SMRTbell Template Prep Kit-SPv3 (Pacific Biosciences, Menlo Park, CA, USA) and sequenced on the PacBio Sequel system. The Illumina sequencing library was constructed using the NEBNext Ultra II DNA Library Prep Kit for Illumina (New England Biolabs, Ipswich, MA, USA) and sequenced on the Illumina HiSeq system.