Early and late disease scleroderma patient and healthy control samples RNA seq
4 mm punch biopsies were obtained from lesional forearm skin and, for a subset of patients, non-lesional back skin from 15 SSc patients ( with disease duration < 2.5 year classified as early stage and disease duration > 8 years  classified as late stage) and 6 healthy volunteers. Total RNA extraction was performed using the Qiagen RNeasy Fibrous Tissue Mini Kit. RNA-seq libraries were generated from 100 ng total RNA prepared using the Illumina TruSeq Stranded Total RNA Library Prep Kit with Ribo-Gold rRNA depletion. Samples were then multiplexed and sequenced on anNextSeq 500 sequencer, producing an average of 80100 million 75-bp paired-end reads per sample.