Instrument: Illumina HiSeq 2000
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: SINGLE
Construction protocol: Total cellular RNA was extracted from ~1-5x10^6 cells using the RNeasy Protect Mini kit or RNeasy Plus kit (Qiagen, Valencia, CA), according to the manufacturer's instructions. For RNA library generation from human and chimpanzee NPCs and eight-week-old neurons, polyA+ RNA was fragmented and prepared into sequencing libraries using the Illumina TruSeq RNA sample preparation kit. NPC-derived sequencing libraries were analyzed on an Illumina HiSeq 2000 sequencer at the UCSD Biomedical Genomics Laboratory (BIOGEM). cDNA libraries were prepared from two human and two chimpanzee NPC lines (two clones each) derived from human WT-33 and ADRC-40 iPSC lines and chimpanzee PR00818 and PR01209 iPSC lines, respectively. Libraries were sequenced using single-end 100 bp reads at a depth of 15-30 million reads per library. Sequencing libraries derived from eight-week-old neurons were analyzed on an Illumina HiSeq 2500 sequencer at the Salk Next Generation Sequencing Core. cDNA libraries were prepared from two human (WT-33 and ADRC-40) and two chimpanzee (PR00818 and PR01209) neurons, one clone each. Libraries were sequenced using paired-end 100 bp reads at a depth of 15-30 million reads per library.