title: HTO_S3
organism: Mus musculus
description: Protocols: FIP200f/fMb1Cre+/- (KO) mice and their littermatecontrol, FIP200f/fMb1Cre-/-(WT) were immunized with 50 ug NP29-KLH with Alum, then splenocytes isolated from WT and KO mice 11 days after immunization were enriched for GC, plasma, and memory B cells by depleting non-B cells and IgDhi B cells using magnetic beads. Those enriched samples were then barcoded and sorted for naïve, GC, plasma, and memory B cells, which were then mixed for single-cell RNA sequencing. Mixed B cells were resuspended in the master mix and loaded together with partitioning oil and gel beads into the chip to generate the gel bead-in-emulsion (GEM). Library was performed according to the manufactor's instruction (Chromium Single-Cell 5′ Reagent Kit v2 Chemistry, Dual Index). Briefly, the poly-A RNA form the cell lysate contained in every signle GEM was retrotranscripted to cDNA, which contains an Illumina R1 primer sequence, the 10x Barcode and Unique Molecular Identifier (UMI). The pooled barcodeed cDNA was then cleaned up with Sliane DynaBeads, amplified by PCR and the apropiated sized fragmnets were selected with SPRIselect reagent for subsequent library constrution. During the library construction, Illumina R2 primer sequence, paired-end constructs with P5 and P7 sequences and Sample index i5 and i7 were added.
attributes.tag: ENA first public
attributes.value: 2025-11-26
attributes.tag: INSDC center name
attributes.value: The Ragon Institute of Mass General Brigham, MIT, and Harvard
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-16123:HTO_S3
attributes.tag: age
attributes.value: 15
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell type
attributes.value: germinal center B cell; naive B cell; Plasma B cell; memory B cell;
attributes.tag: collection date
attributes.value: not collected
attributes.tag: common name
attributes.value: house mouse
attributes.tag: developmental stage
attributes.value: Mature
attributes.tag: genotype
attributes.value: Fip200flox/flox Mb1Cre-/-
attributes.tag: geographic location (country and/or sea)
attributes.value: not collected
attributes.tag: individual
attributes.value: WT
attributes.tag: isolate
attributes.value: not applicable
attributes.tag: organism part
attributes.value: spleen
attributes.tag: sample name
attributes.value: E-MTAB-16123:HTO_S3
attributes.tag: scientific_name
attributes.value: Mus musculus
attributes.tag: sex
attributes.value: female
attributes.tag: strain
attributes.value: Fip200flox/floxMb1Cre-/-