Transcriptomic profiling of bulk tissue and laser-capture microdissected neurons of postmortem human brains in the superior temporal gyrus
Total RNA was extracted using the Direct-zol RNA MiniPrep (Zymo Research #R2051) following manufacturer's protocol, with the inclusion of DNase I treatment and eluted in DNase/RNase-free water. Quality and quantity of RNA were determined via RNA 6000 Nano chip on 2100 Bioanalyzer (Agilent), NanoDrop 2000 spectrophotometer (ThermoFisher Scientific), and Qubit fluorometer (ThermoFisher Scientific).  Fifty ng of RNA were used to create strand-specific total RNA libraries with the NuGEN Ovation Universal RNA-Seq System v2 (NuGEN kit #9102) and processed in parallel on the Sciclone NGS automated workstation (Perkin Elmer) according to manufacturer protocol. Following second strand cDNA synthesis, samples were sheared by sonication on the Covaris E220.  InDA-C (aka AnyDeplete) primers were used to target and cleave adapters from rRNA transcripts before amplification of libraries through 16 cycles of PCR. Final barcoded libraries were bead purified and examined for QC using 2100 BioAnalyzer DNA High Sensitivity chips.  Library concentration was calculated based on fragment size and normalized to 15 nM for sequencing.