title: Sample 3
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 2x10E7 PFU UV-inactivated vesicular stomatitis virus (VSV) on day 0 and boosted with a subsequent s.c. immunization with 5x10E7 PFU UV-inactivated VSV on day 4. UV-inactivation was performed by exposing virus to UV light for 20 minutes. Germinal center B cells from draining inguinal lymph nodes were examined at day 12 following the first VSV 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: 2020-01-13T04:04:20Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2019-11-11T11:09:32Z
attributes.tag: External Id
attributes.value: SAMEA6167457
attributes.tag: INSDC center name
attributes.value: Institute of Immunobiology, Kantonsspital St. Gallen, St. Gallen, Switzerland
attributes.tag: INSDC first public
attributes.value: 2020-01-13T04:04:20Z
attributes.tag: INSDC last update
attributes.value: 2019-11-11T11:09:32Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-8454:Sample 3
attributes.tag: age
attributes.value: 6 to 8
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: clinical information
attributes.value: VSV s.c. immunized
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++
attributes.tag: individual
attributes.value: 4 mice pooled
attributes.tag: infect
attributes.value: vesicular stomatitis virus
attributes.tag: inferred cell type
attributes.value: B-cell
attributes.tag: organism part
attributes.value: lymph node
attributes.tag: sample name
attributes.value: E-MTAB-8454:Sample 3
attributes.tag: scientific_name
attributes.value: Mus musculus
attributes.tag: sex
attributes.value: mixed
attributes.tag: strain
attributes.value: C57Bl/6N