organism: Mus musculus
description: Protocols: The following mouse C57BL/6 (B6) primary cell lines were purchased and cultured for RNA isolation according to manufacturer's instructions. Bone Marrow Mononuclear cells, c57-6271F, Caltag Medsystems, Buckingham, UK; Mouse Embryonic Fibroblasts, untreated, GSC-6002, AMS Biotechnology Ltd; Mesenchymal Stem cells, MUBMX-01001, Cambridge Bioscience, Cambridge, UK; embryonic stem cell line #693 ES C57BL/6 (SCRC-1002, from LGC standards, Middlesex, UK). Embryonic mammary progenitor cell (eMPC) clones were derived from E12.5-stage mammary organs. After microdissection, mammary primordial tissue was cultured on thick basement membrane extract for 4 weeks. After enzymatic dissociation eMPCs were plated and expanded in 2D culture to obtain a pool of eMPCs. eMPC cells were subject to single cell sorting using FACS. Clones were expanded as single-cell derived clones (eMPC1, 2, etc.) to create the eighteen single cell-derived clones described in this study. Cells were harvested by trypsinisation. Total RNA from cells were extracted using RNeasy Mini kit 74104 Qiagen, Manchester, UK. cDNA library preparation was carried out at Oxford Genomics Centre, The Wellcome Trust Centre for Human Genetics using PolyA+ RNA enrichment method for total RNA from cultured cells . mRNA fraction was selected from the total RNA before conversion to cDNA. Second strand cDNA synthesis incorporated dUTP. The cDNA was end-repaired, A-tailed and adapter-ligated. Prior to amplification, samples underwent uridine digestion. The prepared libraries were size selected, multiplexed and quality checked.
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2018-11-30T11:02:52Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2018-06-06T11:53:45Z
attributes.tag: External Id
attributes.value: SAMEA4711897
attributes.tag: INSDC center name
attributes.value: The Institute of Cancer Research
attributes.tag: INSDC first public
attributes.value: 2018-11-30T11:02:52Z
attributes.tag: INSDC last update
attributes.value: 2018-06-06T11:53:45Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-6856:eP.1
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell type
attributes.value: mammary stem cell
attributes.tag: common name
attributes.value: house mouse
attributes.tag: developmental stage
attributes.value: embryonic day 12.5
attributes.tag: genotype
attributes.value: Tg(H2-K1-tsA58)11Kio/s-SHIP-GFP
attributes.tag: organism part
attributes.value: mammary bud
attributes.tag: sample name
attributes.value: E-MTAB-6856:eP.1
attributes.tag: scientific_name
attributes.value: Mus musculus
attributes.tag: sex
attributes.value: female
attributes.tag: strain
attributes.value: (C57BL/6 x (CBA/Ca x C57BL/10)_ F6