title: B091 CD57-
organism: Homo sapiens
description: Protocols: Blood samples were taken from five HLA-A*24+ males with newly diagnosed type 1 diabetes. PBMCs were isolated on density gradients using Lymphoprep (Nycomed) and washed in RPMI supplemented with 2% human AB serum. PBMCs were washed and stained as above and CD57+ and CD57- effector memory CD8 T cells (gated as live CD14-CD16-CD19-CD4-CD3+CD8+CCR7-CD45RA-CD27-) were FACS-sorted using an Aria (BD Biosciences) into BSA supplemented with 10% BSA/5 mM EDTA. RNA was isolated immediately from cell pellets using the PicoPure RNA extraction kit (ThermoFisher). cDNA was generated and amplified using the SMARTseq v4 Ultra Low Input RNA Kit (Clontech) on the contactless Labcyte liquid handling system (Labcyte Inc). 200 ng of amplified cDNA was used from each sample where possible to generate libraries using the Ovation Ultralow Library System V2 kit (NuGEN). In brief, cDNA was fragmented through sonication on Covaris E220 (Covaris Inc), repaired and polished followed by ligation of indexed adapters. Adapter ligated cDNA was pooled before final amplification to add flow cell primers.
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2018-08-03T17:03:02Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2018-04-27T15:17:23Z
attributes.tag: External Id
attributes.value: SAMEA4618641
attributes.tag: INSDC center name
attributes.value: King's College London
attributes.tag: INSDC first public
attributes.value: 2018-08-03T17:03:02Z
attributes.tag: INSDC last update
attributes.value: 2018-04-27T15:17:23Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-6727:B091 CD57-
attributes.tag: age
attributes.value: 35
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell type
attributes.value: CD8-Positive T-Lymphocytes
attributes.tag: common name
attributes.value: human
attributes.tag: disease
attributes.value: type I diabetes mellitus
attributes.tag: individual
attributes.value: B091
attributes.tag: organism part
attributes.value: blood
attributes.tag: phenotype
attributes.value: CD57-
attributes.tag: sample name
attributes.value: E-MTAB-6727:B091 CD57-
attributes.tag: scientific_name
attributes.value: Homo sapiens
attributes.tag: sex
attributes.value: male