LG1_3
Homo sapiens
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).
ENA-FIRST-PUBLIC
2020-07-24T04:03:36Z
ENA-LAST-UPDATE
2019-09-18T17:05:35Z
External Id
SAMEA5785276
INSDC center name
MRC WT Cambridge Stem Cell Institute Department of Surgery
INSDC first public
2020-07-24T04:03:36Z
INSDC last update
2019-09-18T17:05:35Z
INSDC status
public
Submitter Id
E-MTAB-8347:LG1_3
broker name
ArrayExpress
cell cycle phase
Late G1
cell line
H9
cell type
embryonic stem cell
common name
human
developmental stage
embryo
disease
normal
genotype
wild type genotype
organism part
inner cell mass
sample name
E-MTAB-8347:LG1_3
scientific_name
Homo sapiens