alias: BC5YCLACXX A017
title: RNA-Seq of Bos taurus: adult granulosa cells. Non-carrier female A017
design: RNA libraries were generated using the Illumina TruSeq RNA Sample Preparation Kit (Illumina Inc., San Diego, California, USA) per the manufacturer's protocol.  mRNA was purified from 1µg total RNA using poly-T oligo-attached magnetic beads, fragmented using divalent cations under elevated temperature, and purified.  Double-stranded complementary DNAs (cDNAs) were synthesized using SuperScript II Reverse Transcriptase (Invitrogen, Carlsbad, California, USA) and random primers, followed by removal of mRNA using RNAse and purification using Agencourt AMPure XP beads (Qiagen, Valencia, California, USA).   DNA fragments were ligated to Illumina adapters and amplified for 9 cycles using PhusionTM DNA Polymerase and Illumina's PE genomic DNA primer set followed by purification.  Quality and quantity of the finished libraries were assessed using an Agilent DNA1000 series chip assay (Agilent Technologies, Santa Clara, CA) and Invitrogen Qubit HS Kit (Invitrogen, Carlsbad, California, USA), respectively.  Each library was standardized to a 2 _M concentration.  Cluster generation was performed using a TruSeq Single Read Cluster Kit (v4) and the Illumina cBot, with libraries multiplexed for 2 x 100 bp paired-end sequencing using the TruSeq 100 bp SBS kit (v3) and HCS1.6 software, on an Illumina HiSeq2000. All samples were co-loaded on two flow cells.  Images were analyzed using CASAVA (Illumina Inc., San Diego, California, USA).
identifiers.namespace: SUB2923145
identifiers.id: BC5YCLACXX A017