title: Sample 4
organism: Mus musculus
description: Protocols: Draining inguinal lymph nodes were harvested, collected in PBS and gently smashed across a wire mesh. The dissociated cell suspension was stained with anti-CD19, anti-B220, anti-CD38, anti-GL7 and the viability marker 7AAD in PBS containing 0.5% FCS and 10mM EDTA. VSV immunization protocol:Mice were immunized sub-cutaneously (s.c) with 50 micrograms of NP-KLH in alum on day 0. Germinal center B cells from draining inguinal lymph nodes were examined at day 12 following the NP-KLH immunization. CD19+ CD38- GL7+ cells from lymph nodes of immunized Cxcl13-Cre/TdTomato EYFP or Cxcl13-Cre/TdTomato Cxcl12fl/fl EYFP mice were sorted using the BioRad S3 Cell Sorter or the BD FACS Melody cell sorter. Single cell suspensions were run on the 10x Chromium (10x Genomics) system. The cDNA library generation was performed following the established commercial protocol for Chromium Single Cell 3' Reagent Kit (v2 Chemistry). For B cell repertoire analysis, the Chromium Single Cell 5' Library Construction and Mouse V(D)J Enrichment kits (V1.0 Chemistry) were used.
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2020-05-12T17:04:59Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2020-04-07T11:33:08Z
attributes.tag: External Id
attributes.value: SAMEA6650830
attributes.tag: INSDC center name
attributes.value: Institute of Immunobiology, Kantonsspital St. Gallen, St. Gallen, Switzerland
attributes.tag: INSDC first public
attributes.value: 2020-05-12T17:04:59Z
attributes.tag: INSDC last update
attributes.value: 2020-04-07T11:33:08Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-8904:Sample 4
attributes.tag: age
attributes.value: 6 to 8
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell type
attributes.value: germinal center B cell
attributes.tag: common name
attributes.value: house mouse
attributes.tag: developmental stage
attributes.value: adult
attributes.tag: genotype
attributes.value: Tg(Cxcl13-Cre/TdTomato)+ x R26-EYFP++ x Cxcl12fl/fl
attributes.tag: immunophenotype
attributes.value: CD19+ CD38- GL7+
attributes.tag: organism part
attributes.value: inguinal lymph node
attributes.tag: replicate
attributes.value: 2
attributes.tag: sample name
attributes.value: E-MTAB-8904:Sample 4
attributes.tag: scientific_name
attributes.value: Mus musculus
attributes.tag: sex
attributes.value: mixed
attributes.tag: stimulus
attributes.value: subcutaneous NP-KLH immunization
attributes.tag: strain
attributes.value: C57BL/6N