RNA-sequencing of subcutaneous adipose tissue from needle biopsy near umbillicus
We extracted total RNA from the frozen biopsy samples using the Qiagen miRNeasy kit and performed polyA+ selection and library preparation using Illumina TruSeq RNA Sample Preparation Kit v2. We sequenced on an Illumina HiSeq 2000 using 50bp paired-end reads at the University of California Los Angeles Neuroscience Genomics Core. In addition to the fastq files, the molecular data file includes raw counts per gene for each individual. We filtered out adapter sequences using TagDust and required sequencing reads to have a Phred quality score > 20 using Fastx-toolkit. We aligned the RNA-Seq reads using STAR (v.2.4.2a) 2-pass alignment and the GENCODE v19 annotation.