RNA-seq of Human Fibroblasts: Control rep 1
RNA was extracted using Bio-Rad Aurum kit from fibroblasts. RNA quality was checked and the NEBNext Ultra II Directional RNA Library Prep Kit for Illumina with NEBNext rRNA Depletion Kit was used to prepare RNA-seq libraries, with a total of 500 ng input RNA from each sample. The manufacturers protocols were followed, with the following exceptions: 40X adaptor dilutions used, all bead incubations done at room temp, used 4X lower concentrations of index primers, and ten cycles of library amplification were performed. The resulting libraries were pooled in equimolar amounts, quantified using the KAPA Library Quant Kit for Illumina, quality checked, and were sequenced using paired-end, 75 base pair sequencing on the Illumina NextSeq 500 massively parallel sequencer at the University of Florida Center for NeuroGenetics.