title: UPN4_j10
organism: Homo sapiens
description: Protocols: Whole human peripheral blood was collected into sterile vacutainer tubes containing EDTA for single cell RNA sequencing, tubes were centrifuged at 300 g for 5 min at room temperature and plasma was collected. To fully capture peripheral blood cell heterogeneity, we analyzed fresh samples without cell sorting or freezing and without Ficoll enrichment, minimizing time of incubation and processing. Sample preparation was done at room temperature. After red cell lysis, single-cell suspensions were loaded onto a Chromium Single Cell Chip (10x Genomics) according to the manufacturer's instructions for co-encapsulation with barcoded Gel Beads at a target capture rate of ~7000 individual cells per sample. To analyze neutrophils, we added RNase inhibitor (RNAse OUT Recombinant Ribonuclease Inhibitor Invitrogen, 40U/mL) into the loading buffer. Captured mRNAs were barcoded during cDNA synthesis using the Chromium Single Cell 3' Solution v3 (10x Genomics) according to the manufacturer's instructions. Of note, we increased the PCR cycles by two during cDNA amplification. All samples (at Day 0 and Day 10) were processed simultaneously with the Chromium Controller (10x Genomics) and the resulting libraries were prepared in parallel in a single batch.
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2020-08-22T04:06:45Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2020-06-18T05:03:27Z
attributes.tag: External Id
attributes.value: SAMEA6979317
attributes.tag: INSDC center name
attributes.value: Inserm U1015, Gustave Roussy Cancer Cancer Villejuif, France
attributes.tag: INSDC first public
attributes.value: 2020-08-22T04:06:45Z
attributes.tag: INSDC last update
attributes.value: 2020-06-18T05:03:27Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-9221:UPN4_j10
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: common name
attributes.value: human
attributes.tag: disease
attributes.value: COVID-19
attributes.tag: individual
attributes.value: UPN4
attributes.tag: organism part
attributes.value: blood
attributes.tag: phenotype
attributes.value: mild COVID-19
attributes.tag: sample name
attributes.value: E-MTAB-9221:UPN4_j10
attributes.tag: scientific_name
attributes.value: Homo sapiens