title: lane1
organism: Homo sapiens
description: Protocols: MKs were obtained by culture of CD34+ hematopoietic progenitor cells. In short, cord blood was collected with informed consent and CD34+ cells were prepared by magnetic bead selection. Purified cells were cultured (1x105 cells/ml) for 10 days in serum-free medium supplemented with human thrombopoietin and interleukin-1beta to differentiate into MKs.By day 10 more than 90% of cells stained positive for CD41 positive and negative for CD34. Total RNA was obtained by the Trizol method. Polyadenylated RNA was purified from this by two rounds of magnetic Oligo dT25 selection (Dynabeads Oligo (dT)25, Invitrogen) in the presence of RNase inhibitor (New England Biolabs, Hitchin, UK), combined with DNase treatment (DNA-free, Ambion, Invitrogen). The mRNA was fragmented using RNA fragmentation reagent (Ambion, Invitrogen) and precipitated with added glycogen (Ambion, Invitrogen), then cDNA produced by Superscript Double Stranded cDNA synthesis kit (Invitrogen), replacing oligo-dT primer with random primer. Thereafter, library preparation was as per mRNA-sequencing kit (Illumina, Little Chesterford, UK), starting with end repair.
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2012-01-09T14:41:02Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2018-03-08T15:35:14Z
attributes.tag: External Id
attributes.value: SAMEA1094720
attributes.tag: INSDC center name
attributes.value: SC
attributes.tag: INSDC first public
attributes.value: 2012-01-09T14:41:02Z
attributes.tag: INSDC last update
attributes.value: 2018-03-08T15:35:14Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-918:lane1
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell type
attributes.value: megakaryocyte
attributes.tag: common name
attributes.value: human
attributes.tag: organism part
attributes.value: cord blood
attributes.tag: sample name
attributes.value: E-MTAB-918:lane1
attributes.tag: scientific_name
attributes.value: Homo sapiens