title: 1A
organism: Homo sapiens
description: Protocols: The local Regional Ethical Review Board (Link_ping, Sweden) granted an ethical permission for this study. Unstimulated non-irradiated apheresis platelets were collected from two healthy human male and one healthy female blood donor. Platelets were collected by COBE Spectra system (Gambro BCT) and were used on the same collection day. Residual leukocytes were depleted with anti-CD45 conjugated Dynabeads, according to the manufacturers recommendations (Pan Leukocyte; Invitrogen, Carlsbad, CA). The platelet suspension, with a volume of 30 mL and a platelet count of 1.4 * 10exp9 cells/mL, was centrifuged at 800 g for 8 min and the supernatant was discarded. The platelet pellet was processed for leukocyte depletion, as recommended by the supplier of Pan Leukocyte reagent (Invitrogen, Carlsbad, CA). The leukocyte removal was performed at room temperature. Approximately 70 _ 75% of the original platelets were recovered after the leukocyte depletion. To investigate the level of leukocyte contamination, we determined the level of CD45 (PTPRC) transcript by qPCR using TaqMan Gene Expression Assay for this gene according to the recommendations of the supplier (Assay ID: Hs00894713_m1; Applied Bioystems, Carlsbad, CA, USA). For isolation of total RNA, we employed the miRVana RNA Extraction Kit as recommended by the supplier (Life Technologies). Ribosomal RNA was depleted with Ribo-Zero (Epicentre, Madison, WI, USA) and strand specific barcoded RNA-sequencing libraries were prepared using ScriptSeq v2 (Epicentre) according to manufacturers instructions.
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2013-10-01T17:01:24Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2018-03-08T16:41:44Z
attributes.tag: External Id
attributes.value: SAMEA2175888
attributes.tag: INSDC center name
attributes.value: Dept of Clinical and Experimental Medicine, Linksping University, Sweden
attributes.tag: INSDC first public
attributes.value: 2013-10-01T17:01:24Z
attributes.tag: INSDC last update
attributes.value: 2018-03-08T16:41:44Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-1846:1A
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell type
attributes.value: platelet
attributes.tag: common name
attributes.value: human
attributes.tag: individual
attributes.value: RNA Extract S1
attributes.tag: sample name
attributes.value: E-MTAB-1846:1A
attributes.tag: scientific_name
attributes.value: Homo sapiens
attributes.tag: sex
attributes.value: male
attributes.tag: specimen with known storage state
attributes.value: fresh specimen