Transcriptomic profiling of bulk tissue and laser-capture microdissected neurons of postmortem human brains in the superior temporal gyrus
Using a Leica LMD-6000 laser capture microdissection system, 100 neurons from each sample were laser captured directly into lysis buffer (PicoPure RNA Isolation Kit, ThermoFisher Scientific KIT0204). RNA was extracted using the PicoPure total RNA kit with inclusion of DNase I digest according to manufacturer protocol. Strand-specific rRNA depleted RNA libraries were prepared from 10 μl of the final neuronal RNA eluate using the NuGEN Ovation SoLo Kit (NuGEN Kit #0500) for ultra-low input following manufacturer protocol with final amplification of 18 PCR cycles.  Barcoded bead purified libraries were examined for QC using Qubit Fluorometer (ThermoFisher Scientific) and 2100 BioAnalyzer DNA High Sensitivity chips.  Library concentration was calculated based on fragment size and normalized to 15 nM for sequencing. For sequencing, a custom R1 primer from the SoLo kit (NuGEN) was used in place of the standard Illumina forward read primer.