title: GSM4783645: snRNAseq_AR6_Gfap-Cre_Wlsfl+_RP2; Mus musculus; ncRNA-Seq
library_construction_protocol: both hippocampi of a single male mouse were rapidly dissected in ice-cold homogenization buffer (0.25 M sucrose, 25 mM KCl, 5 mM MgCl2, 20 mM Tricine-KOH).  The tissue was minced with a razor blade and Dounce homogenized using a loose-fitting pestle in 5 ml of homogenization buffer supplemented with 1 mM DTT, 0.15 mM spermine, 0.5 mM spermidine, and EDTA-free protease inhibitor (Roche 11 836 170 001), and 60 U/mL RNasin Plus RNase Inhibitor (Promega N2611).  A 5% IGEPAL-630 solution was added to bring the homogenate to 0.3% IGEPAL-630, and the homogenate was further homogenized with five strokes of a tight-fitting pestle.  The sample was filtered through a 50 μm filter (CellTrix, Sysmex, 04-004-2327), underlayed with solutions of 30% and 40% iodixanol in homogenization buffer (Sigma D1556), and centrifuged at 10,000 x g for 18 minutes in a swinging bucket centrifuge at 4°C.  Nuclei were collected at the 30%-40% interface, diluted with 2 volumes of homogenization buffer and concentrated by centrifugation for 10 min at 0.5 x g at 4°C.  snRNA-seq sequencing libraries were constructed using the 10X Genomics Chromium single cell 3' v3 kit.