title: GSM9253511: Microglia, WT, Basal, 1; Homo sapiens; RNA-Seq
library_construction_protocol: In order to obtain RNA from both the adherent MGLs and the MGLs suspended in culture, the media from 28-day, mature MGLs in a 6-well plate were collected, centrifuged for 5 min at 300 x g, washed once with PBS and recentrifuged for 3 min at 500 x g. Adherent MGLs from the same plate were washed once with PBS. Then RNA was isolated directly from the well using Arcturus PicoPure RNA Isolation Kit (Thermo Fisher Scientific 2970541) according to manufacturer's instructions. The adherent and nonadherent microglia were combined in RNA extraction buffer. One well per line was collected over four batches of differentiation. RNA libraries were prepared from 100 nanograms of total RNA using Illumina Stranded mRNA Prep, Ligation kit (Illumina), following the manufacturer's instructions.