description: Protocols: Peripheral blood mononuclear cells (PBMC) from the year 2010 (Donor1) and year 2014 (Donor11) were isolated by the standard density-gradient centrifugation and cryopreserved until the day of use. Blood samples collected in 2020 from both Donor1 and Donor2 were processed immediately to isolate memory B cells and plasma cells by flow cytometry followed by single-cell immune profiling using the 10X system. Single cell suspension was performed according to Chromium Next GEM Single cell V(D)J Reagent kits v1.1. The positive fractions of memory B cells and plasma cells were collected in 6ul of FCS containing eppendorf tubes that were pre-coated overnight with 2percent BSA. The 5'scRNA-seq libraries were generated using Chromium Next GEM Single Cell V(D)J Reagent Kit v1.1 or v2 (10X Genomics) according to the manufacturer's protocol. Paired heavy and light chain BCR libraries were prepared from the sorted B cell populations. All the steps were followed as per the manufacturer's instructions in user guide recommended for 10X Genomics kit v1.1 or 2.
attributes.tag: ENA first public
attributes.value: 2022-04-26
attributes.tag: ENA last update
attributes.value: 2022-04-26
attributes.tag: External Id
attributes.value: SAMEA11308716
attributes.tag: INSDC center alias
attributes.value: Istituto Nazionale Genetica Molecolare (INGM)
attributes.tag: INSDC center name
attributes.value: Istituto Nazionale Genetica Molecolare (INGM)
attributes.tag: INSDC first public
attributes.value: 2022-04-26T16:19:20Z
attributes.tag: INSDC last update
attributes.value: 2022-04-26T16:19:20Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-11174:Donor2_PC_2014
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell type
attributes.value: plasma cell
attributes.tag: common name
attributes.value: human
attributes.tag: developmental stage
attributes.value: adult
attributes.tag: disease
attributes.value: normal
attributes.tag: individual
attributes.value: 2
attributes.tag: organism part
attributes.value: blood
attributes.tag: sample name
attributes.value: E-MTAB-11174:Donor2_PC_2014