title: GSM2695603: OHara_sample_72; Eumetopias jubatus; RNA-Seq
library_construction_protocol: Whole blood collected in PaxGene™ RNA tubes were placed in a cooler with ice until transport back to the ship, at which point they were kept in the freezer at approximately -15°C. Upon return to UAF the samples were placed in a -80°C freezer and then shipped to the University of Washington Center for Exposures, Diseases, Genomics and Environment in insulated shipping containers with dry ice and subsequently stored at -80°C until extraction. Total RNA was extracted using the PaxGene™ Blood RNA Kit according to the manufacturer’s protocol. RNA integrity was determined using an Agilent 2100 Bioanalyzer (Agilent Technologies, Palo Alto, California).  RNA quantity was determined by measuring OD260 with a Thermo Scientific NanoDrop™ 1000 Spectrophotometer (Thermo Fisher Scientific, Inc.; Wilmington, DE). RNA samples were depleted of ribosomal RNA using the Low Input RiboMinus Eukaryotic System v2 kit (Thermo Fisher Scientific). The cDNA library was then created with the ribosomal depleted RNA using the Ion Total RNA-Seq Kit v2 (Thermo Fisher Scientific). This included fragmentation via RNase III, hybridizing and ligating the RNA, performing reverse transcription, and amplification.  Samples were labeled with the Ion Xpress RNA-Seq Barcode 1-16 Kit (Thermo Fisher Scientific) to enable multiplexing 4 samples per run.  Templating, loading and sequencing were carried out using the Ion PI Template OT2 200 Kit v2, the Ion PI Chip Kit v2, and the Ion PI Sequencing 200 Kit v2 respectively (all from Thermo Fisher Scientific).
attributes.tag: GEO Accession
attributes.value: GSM2695603