RNA-Seq of homo sapiens: adult female Muscle - Skeletal
Total RNA was quantified using the Quant-iT RiboGreen RNA Assay Kit and normalized to 5 ng per uL. An aliquot of 200 ng was transferred into library preparation, which was an automated variant of the Illumina TruSeq protocol (Revision A, 2010; based on polyA selection and not strand-specific) with Broad Institute-designed indexed adapters substituted in for multiplexing. The libraries were quantified with qPCR using the KAPA Library Quantification Kit and then pooled equimolarly. Pooled libraries were normalized to 2 nM and denatured using 0.1 N NaOH prior to sequencing. Flow cell cluster amplification and sequencing were performed according to the manufacturer's protocols using either the HiSeq 2000 or HiSeq 2500.