organism: Homo sapiens
description: Protocols: FUCCI hESCs were washed with PBS and collected by incubating with Cell Dissociation Buffer (Gibco) for 10 min at 37°C. Cells were washed with 1% PBS/BSA and incubated in 1% PBS/BSA with anti-TRA-1-60-antibody mouse monoclonal IgM (1:100, Santa Cruz Biotechnology, Inc.) for 20 min on ice, together with the secondary antibody, Alexa Fluor® 647 goat anti-mouse IgM (1:1,000, Thermo Fisher). Cells were washed with PBS, re-suspended in 6ml of PBS and sorted into the different cell cycle phases using BD FACS AriaTM III, BD Fusion or BD Influx sorters by the Cambridge NIHR BRC Cell Phenotyping Hub. RNA was extracted using GenEluteTM Mammalian Total RNA Miniprep Kit and On-Column DNase I Digestion Set (Sigma Aldrich) according to manufacturer instructions. Polyadenylated (poly-A) purified opposing strand-specific mRNA library libraries were prepared from 1μg of total RNA using the TruSeq Stranded mRNA sample preparation kit (Illumina).
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2020-07-24T04:03:36Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2019-09-18T17:05:35Z
attributes.tag: External Id
attributes.value: SAMEA5785275
attributes.tag: INSDC center name
attributes.value: MRC WT Cambridge Stem Cell Institute Department of Surgery
attributes.tag: INSDC first public
attributes.value: 2020-07-24T04:03:36Z
attributes.tag: INSDC last update
attributes.value: 2019-09-18T17:05:35Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-8347:LG1_2
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell cycle phase
attributes.value: Late G1
attributes.tag: cell line
attributes.value: H9
attributes.tag: cell type
attributes.value: embryonic stem cell
attributes.tag: common name
attributes.value: human
attributes.tag: developmental stage
attributes.value: embryo
attributes.tag: disease
attributes.value: normal
attributes.tag: genotype
attributes.value: wild type genotype
attributes.tag: organism part
attributes.value: inner cell mass
attributes.tag: sample name
attributes.value: E-MTAB-8347:LG1_2
attributes.tag: scientific_name
attributes.value: Homo sapiens