GSM8078518: Control, C053; Bos taurus; RNA-Seq
Peripheral blood was sampled from each animal using blood collection tubes with blood harvested from the tail vein.  All tubes were inverted 4-6 times immediately after sampling and transported to the laboratory in a refrigerated cool box within three hours of collection.  Whole blood was collected in Tempus™ blood RNA stabilisation tubes (Thermo Fisher Scientific) for isolation of total RNA.  All tubes intended for RNA extraction were stored at −80°C prior to isolation and purification. Total RNA preparation was performed for each animal (bTB+ and bTB−) using the Tempus™ Spin RNA Isolation Kit (Thermo Fisher Scientific). The Tempus™ tubes were inverted to mix the contents, which were then poured into sterile 50 ml conical tubes and total RNA was isolated and purified according to the manufacturer's instructions. Following total RNA purification, an Agilent 2100 Bioanalyzer and the RNA 6000 Nano LabChip kit (Agilent Technologies, Inc.) were used to quality check and generate RNA integrity number (RIN) values for all samples, which were then stored at −80°C. Total RNA samples (bTB+ and bTB−) were used with the NEBNext® Ultra™ RNA Library Prep Kit for Illumina® (New England Biolabs) to construct sequence barcode-indexed RNA-seq libraries, which were then used for 150 bp paired-end sequencing on the NovaSeq™ 6000 Sequencing System (Illumina, Inc.). RNA-seq library preparation and sequencing were performed by an external provider (Novogene).