A cold-specialized icefish species underwent major genetic changes as it migrated to temperate waters, new study finds UofIllinois
). Cell density was assessed by counting with a hemacytometer, and appropriate volumes of the hepatocyte suspension were embedded in 1% agarose blocks using disposable plug molds . Hepatocytes were thoroughly lysed in-agarose using 1% lithium dodecyl sulfate in notothenioid ringer . The blocks were then fully equilibrated with a preservation buffer that permitted return transport at ambient temperature, and were subsequently kept at 4 °C until use.
Specimen collection and sampling were conducted with permits granted by the Chilean Fisheries Service under Technical Memorandum P.INV N° 244-2016, the Government of South Georgia and the South Sandwich Islands Regulated Activity Permit 2016-026, and the University of Illinois at Urbana-Champaign IACUC approved protocols 07053 and 17148, for the respective field collection and sampling effort.
Pacific Biosciences CLR library preparation and sequencing were carried out at the University of Oregon Genomics and Cell Characterization Core Facility. The HMW DNA was lightly sheared with Megaruptor at 60 kb target length for library construction using PacBio SMRTbell Express Template Prep kit 2.0. The resulting library was selected for inserts approximately >30 kb with the BluePippin and sequenced on two SMRT cells 8M for each species on Sequel II for 30 h of data capture. A total of 12.
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