Protocols: Blood samples were collected in sodium heparin vacutainer tubes (Becton Dickinson) and peripheral blood mononuclear cells (PBMC) were isolated by Ficoll-plaque (GE Healthcare). PBMC were cryopreserved in 10% DMSO 90% fetal calf serum until analysis. PBMCs were thawed and cultured overnight in RPMI containing 10% human AB serum (Sigma) with 10 U/ml recombinant IL-2. PBMC from the donor TSF were stained with HLA-E/VMAPRTLIL tetramer-PE and CD8β-PECy5. Single tetramer+CD8β+ cells at high or low tetramer staining intensity were sorted directly into individual wells of separate 96-well plates (Eppendorf) using a FACSAria I (Becton Dickinson) at 80 cells per plate into a dNTP mix and lysis buffer. Libraries were prepared with the modified Smart-seq2 protocol essentially as described in (Picelli, S. et al. Full-length RNA-seq from single cells using Smart-seq2. Nat Protoc 9, 171-181 (2014)). Nextera XT DNA sample preparation kit and Index kit A and D (Illumina).
ENA first public
2021-05-21
ENA last update
2021-05-21
External Id
SAMEA8572696
INSDC center alias
University of Melbourne at the Peter Doherty Institute for Infection and Immunity
INSDC center name
University of Melbourne at the Peter Doherty Institute for Infection and Immunity
INSDC first public
2021-05-21T00:12:37Z
INSDC last update
2021-05-21T00:12:37Z
INSDC status
public
Submitter Id
E-MTAB-10305:TSF TRBV14 negative 44
age
49
broker name
ArrayExpress
cell type
T cell
common name
human
developmental stage
adult
disease
normal
immunophenotype
T cell receptor beta variable 14 negative
organism part
blood
sample name
E-MTAB-10305:TSF TRBV14 negative 44
sex
male
single cell quality
OK