alias: GSM4872467
title: GSM4872467: GFP-positive tanycytes P45 Ctrl (scRNA-seq); Mus musculus; RNA-Seq
library_construction_protocol: 2 mm thick coronal slices including the hypothalamic protruding median eminence (ME) were collected using adult mouse brain matrix (Kent Scientific).  The mediobasal hypothalamic region was microdissected using a surgical scalpel, dampened in Hibernate-A media supplemented with 0.5 mM GlutaMax and 2% B-27 (HABG) and chopped with a razor blade. Brain tissues were transferred into pre-equilibrated papain/Dnase-I mix (Worthington Papain Dissociation System, #LK003150) and incubated for 30 min at 37°C with frequent agitation using a fire-polished glass pipette.  Dissociated cells were filtered through a 40 μm strainer and subjected to density gradient centrifugation to remove the cell debris as suggested in the manufacturer's protocol. Cells were resuspended cells with ice-cold PBS containing 0.04% BSA and 0.5 U/μl RNase inhibitor, and 10,000-15,000 cells were loaded on a 10x Genomics Chromium Single Cell system (10X Genomics, Redwood City, CA), using the v3 chemistry following the manufacturer's instructions, and libraries were sequenced on an Illumina NextSeq with ~200 million reads per library.  Sequencing results were processed through the Cell Ranger 3.1 pipeline (10x Genomics, Redwood City, CA) with default parameters. single cell RNAseq
identifiers.namespace: GEO
identifiers.id: GSM4872467
attributes.tag: GEO Accession
attributes.value: GSM4872467