title: PacBio RS II sinlge-end Sequencing of KAS22054414 (KAE22249800)
design: Construction of reference genome of 'Baekdansim' and development of cold resistance markers
library_name: white_Hibiscus_syriacus_PACBIO_GENOMIC
library_construction_protocol: Genomic DNA gDNA of high quality and high molecular weight was extracted from H. syriacus Baekdansim using a protocol described in a previous study1. PacBio genomic DNA library prep was performed according to the manufacturer's instructions https://www.pacb.com/support/documentation/. Purified gDNA was used for library construction by the SMRTbell xpress Template Prep Kit Pacific Biosciences, Cat. No. 101-357-000. In brief, gDNA was mechanically sheared to an average size of 20 kb using a Covaris g-TUBE device Part No. 520079. In total, 5 microgram sheared gDNA was damage-repaired and end-repaired using polishing enzymes. Blunt-end adapter ligation was used to create the SMRTbell emplate. Adapter dimers and contaminants were removed using the AMPure XP bead purification system Beckman Coulter, Cat. No. A63882. A BluePippin size selection system Sage Science, Cat. No. BLU0001 was used to size select the SMRTbell emplate and enrich for fragments >15 kb. Sequencing primer v4 was annealed to the SMRTbell emplate, and DNA polymerase/template complex was created using the Sequel Binding Kit 2.1 Pacific Biosciences, Cat. No. 101-365-900. An additional AMPure XP purification step was performed to remove excess primer and polymerase prior to sequencing. The library was sequenced on a Sequel instrument using SMRT Cell 1M v2 Pacific Biosciences, taking one movie of 10 hours per cell with the Sequel Sequencing Kit 2.0 Pacific Biosciences.
identifiers.namespace: KOBIC
identifiers.id: KAE22249800
identifiers.namespace: Korea Research Institute of Bioscience and Biotechnology
identifiers.id: DRX652878