title: GSM3765247: PP_InVivo_2; Bos taurus; RNA-Seq
library_construction_protocol: Total RNA from each sample was isolated using the RNeasy Micro Kit (Qiagen, Germantown, MD, USA) according to the manufacture's cell protocol with the following adjustments for samples maintained in RNAlater; 250 µl of 100% ethanol was used instead of the recommended 350 µL of 70% (v/v) ethanol and isolated RNA was eluted in 12 µl RNase-free water instead of the recommended 22 µl RNase-free water. Barcoded fragment libraries were constructed using the Bovine Custom Any Deplete Ovation SoLo RNA-Seq System from NuGEN (San Carlos, CA, USA) following the manufacturer's protocol. DNA integrity of the libraries was assessed using the High Sensitivity D1000 ScreenTape on the Agilent 4200 TapeStation (Santa Clara, CA, USA) and DNA concentration was measured using the KAPA qPCR kit (Boston, MA, USA) following the manufacturer's protocol. Samples were then multiplexed and sequenced with 1×75bp single-end reads on an Illumina NEXT-seq (Illumina, Inc.San Diego, CA, USA) at the Stanford Functional Genomics Facility (Stanford, CA, USA) according to the manufacturer's protocol.