title: GSM2794431: F4654; Homo sapiens; RNA-Seq
library_construction_protocol: Fibroblasts were harvested by trypsinization, centrifuged, and rinsed once with ice-cold cold phosphate-buffered saline (PBS).  Total RNA was isolated using a RNeasy Mini Kit (Qiagen) followed by an independent DNase I treatment using the TURBO DNA-free system (Ambion) according to the manufacturer’s recommended rigorous protocol.  RNAs isolated from all 35 fibroblast lines (1 ug/sample) were reverse transcribed to generate sequencing libraries using a Truseq Stranded Total RNA Library Prep kit (Illumina).  This protocol included ribosomal RNA depletion instead of polyA selection.  The cDNA synthesis was conducted using SuperScript II reverse transcriptase followed by second strand synthesis, adenylation of 3’ ends and ligation of adapters. The libraries were sequenced using a 2x76 bases paired end protocol on an Illumina HiSeq 2000 instrument. Totally 35 libraries (18 from FRDA patients and 17 from unaffected controls) were sequenced in 6 lanes, generating 27-39 million pairs of reads per sample. Each pair of reads represents a cDNA fragment from the library. 