organism: Mus musculus
description: Protocols: Mouse spleen isolated, physically disrupted with ground glass, filtered through 70uM mesh, erythrocytes lysed via red blood cell lysis buffer, filtered through 70uM mesh. Cell suspensions were evaluated by light microscopy for debris and viability, and were counted using a hemocytometer. Cell concentrations were 932 and 888 live cells / ul respectfully. Targeting approximately 2400 single cells per sample, single cells were captured, lysed, and RNA was reverse transcribed and barcoded using a 10x Genomics Chromium instrument and Chromium Single Cell 3' Reagent Kits v2 reagents. (10x Genomics, Pleasonton, CA). To construct Illumina compatible sequencing libraries the cDNA is fragmented, A-tail repaired and a double-sided bead cleanup is performed. Adapters are ligated to the cDNA fragments and the fragments are PCR amplified using unique sample index primers per manufacturer's recommendations. Libaries were quantified by qPCR using the KAPA Library Quant Kit (Illumina) from KAPA Biosystems (Roche, Pleasonton, CA).
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2019-03-29T17:02:10Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2019-03-19T11:04:27Z
attributes.tag: External Id
attributes.value: SAMEA5522502
attributes.tag: INSDC center name
attributes.value: UNIVERSITY OF NEBRASKA MEDICAL CENTER
attributes.tag: INSDC first public
attributes.value: 2019-03-29T17:02:10Z
attributes.tag: INSDC last update
attributes.value: 2019-03-19T11:04:27Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-7780:Sample 2
attributes.tag: age
attributes.value: 10
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell type
attributes.value: leukocyte
attributes.tag: common name
attributes.value: house mouse
attributes.tag: developmental stage
attributes.value: adult
attributes.tag: environmental stress
attributes.value: social defeat stress
attributes.tag: genotype
attributes.value: wild type genotype
attributes.tag: individual
attributes.value: 2
attributes.tag: organism part
attributes.value: spleen
attributes.tag: sample name
attributes.value: E-MTAB-7780:Sample 2
attributes.tag: scientific_name
attributes.value: Mus musculus
attributes.tag: sex
attributes.value: male
attributes.tag: strain
attributes.value: C57BL6J