GSM5550060: WT_S2_L002; Mus musculus; RNA-Seq
The hippocampus was dissected from each mouse brain, minced into small pieces, enzymatically dissociated (using papain ~20 units/ml) and mechanically dissociated to generate a single-cell suspension. Myelin was removed using myelin removal beads (Miltenyi Biotec) Libraries were prepared using a Chromium controller according to the 10x Chromium Next GEM Single Cell 3' v3.1 protocol (CG000204). Briefly, cell suspensions were diluted in nuclease-free water to achieve a targeted cell count of 10,000. The cell suspension was mixed with Master Mix and loaded with Single Cell 3'V3.1 Gel Beads and partitioning oil into Chromium Next GEM chip G. RNA transcripts from single cells were uniquely barcoded and reverse-transcribed within droplets. cDNA molecules were pooled, and the cDNA pool was subjected to an end-repair process, the addition of a single 'A' base, and ligation of adapters. The products were then purified and enriched by PCR to create the final cDNA library.