title: RNA-Seq of Homo Sapiens: single brain cells: Sample SC_USC_2619
design: "Single cell RNA was harvested, subjected to aRNA amplification using oligo-dT primers containing T7 promoters as previously described (J Vis Exp. 2011, 50: 2634), with minor modifications. Specifically, we reduced the volume of reactions two times as compared to the method described in the article, and used bead purification instead of ethanol precipitation and RNA column purification. Sequencing library was prepared from 150-400ng aRNA using the TruSeq Stranded mRNA Sample Preparation Kit (illumina, CA, USA) according to manufacturer's protocol. Briefly, cDNA was synthesized using reverse transcriptase (SuperScript III) and random primers. The cDNA was further converted into double stranded DNA, blunt-ended and adenylated at 3'-end. The resulting dsDNA fragments were ligated to double stranded adapters and enriched with PCR using illumina primers. After cleaning up using AMPure XP beads (Beckman, CA, USA), quality control was performed on the resulting library using an Agilent Technologies 2200 TapeStation to characterize DNA fragment size and concentration. The library was subjected to Illumina sequencing platforms on Hiseq2000 and Hiseq2500. Reads were then demultiplexed by index sorting."