description: Protocols: Spleens were recovered immediately after euthanasia, perfused with complete cold medium and cut in small pieces to be crushed on a 70µm filter. Cold ACK was used for red blood cell lysis and cells were kept on ice in medium or complete medium. For surface staining, cells were incubated 30mins on ice with antibodies. Dark zone germinal center B cells sorted as DAPI-CD19+ GL7+IgD-CXCR4highCD86low RNEasy (qiagen) Kit manufacturer protocol. RNA quality was assessed on a Fragment Analyzer (Agilent Technologies). The RNAs had RQNs between 7.6 and 9.4. From 50 - 75 ng total RNA, mRNA was isolated with the NEBNext Poly(A) mRNA Magnetic Isolation Module. RNA-seq libraries were then prepared from the mRNA using the NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (New England Biolabs). Libraries were quantified by a fluorimetric method and their quality assessed on a Fragment Analyzer (Agilent Technologies).
attributes.tag: ENA first public
attributes.value: 2021-02-06
attributes.tag: ENA last update
attributes.value: 2021-01-27
attributes.tag: External Id
attributes.value: SAMEA7875524
attributes.tag: INSDC center alias
attributes.value: CHUV | Service d'immunologie et allergie-IAL
attributes.tag: INSDC center name
attributes.value: CHUV | Service d'immunologie et allergie-IAL
attributes.tag: INSDC first public
attributes.value: 2021-02-06T04:08:54Z
attributes.tag: INSDC last update
attributes.value: 2021-01-27T18:17:21Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-10056:KO1110
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: Was knockout
attributes.tag: organism part
attributes.value: spleen
attributes.tag: phenotype
attributes.value: DAPI-CD19+ GL7+IgD-CXCR4highCD86low
attributes.tag: sample name
attributes.value: E-MTAB-10056:KO1110
attributes.tag: strain
attributes.value: C57BL/6