Leishmania donovani 1S Pacbio Sequel ii reads
Leishmania cells were washed and resuspended in cold PBS buffer prior to lysis and DNAextraction with Nanobind CBB kit (Pacific Biosciences, Menlo Park, CA), following manufacturers protocol for Extracting HMW DNA from cultured adherent cells. Seven micrograms of genomic DNA sample were sheared with g-TUBE (Covaris, Woburn,MA), at 15kb setting. Five micrograms of sheared genomic DNA were prepared into a PacBio sequencing library using SMRTbell Express Template Prep Kit 2.0 (PacificBiosciences, Menlo Park, CA) following manufacturers instructions. After library preparation, the library was size-selected on the BluePippin instrument (Sage Science,Beverly, MA) to remove library fragments smaller than 10kb. Prior to sequencing, the library was bound to sequencing polymerase with Sequel II Binding kit 2.2, then sequenced withSequel II Sequencing kit 2.0 and SMRT cell 8M on the Sequel II instrument (PacificBiosciences, Menlo Park, CA)