RNAseq of n elephant seal blubber during repeated ACTH challenge
cDNA library preparation and Illumina HiSeq4000 sequencing were conducted at the University of California, Berkeley QB3 Vincent J. Coates Genomics Sequencing Laboratory. Ribosomal RNA (rRNA) was depleted using Ribo-Zero rRNA Removal Kit (Human/Mouse/Rat; Illumina, Inc., USA). Yield and quality of mRNA were assessed using fluorometric methods and an Agilent 2100 Bioanalyzer, respectively (Agilent Technologies, USA). PrepX directional RNAseq library kits (WaferGen  Bio-systems Inc., USA) were used for library preparation and libraries were quantified with KAPA Library Quantification kits for Illumina platforms (Kapa Biosystems, USA) using a Bio-Rad CFX Connect (Bio-Rad Laboratories, USA).  Thirteen cycles of indexing PCR using KAPA High Fidelity Hotstart Amplification Kits (Kapa Biosystems, USA) were used before library quantification and validation. Individual libraries were barcoded, indexed as TruSeq single index per library, pooled in equimolar concentrations (400 ng), and sequenced as 100 base-pair paired-end reads on the Illumina HiSeq 4000.