Total RNA-Seq of Homo sapiens: substantial nigra dopamine neuron (SNDA): Sample HC_BC000074_SNDA_3_rep1
Total RNA was isolated and treated with DNase using the Arcturus Picopure method, yielding approximately 7-8 ng RNA per sample. Total RNA was linearly amplified into 5~10 µg of double-stranded cDNA using the validated, precise, isothermal RNA amplification method implemented in the Ovation RNA-seq System V2 (NuGen). Sequencing libraries were generated from 500 ng of the double-stranded (ds) cDNA using the TruSeq RNA Library Prep Kit v2 (Illumina) according to the manufacturer's protocol. The cDNA was fragmented, and end repair, A-tailing, adapter ligation were performed. Library quality and quantity control was performed using the Agilent DNA High Sensitivity Chip and qPCR quantification, respectively. Libraries were sequenced (50 or 75 cycles, paired-end) on an Illumina HiSeq 2500.