Homo sapiens
Protocols: GNS and NS cells were cultured using serum-free basal media supplemented with B27 and N2 culture supplements (Life Technologies). Growth factors EGF and FGF-2 (20 ng/ml) were added. Culture vessels were coated with Laminin (Sigma) for 3 hr at 10 ug/ml prior to use. Alternatively media was directly supplemented with 1 ug/ml Laminin. GNS cells were routinely grown to confluence, dissociated using Accutase (Sigma), and then split 1:3 to 1:7. Medium was replaced every 3-7 days. GNS and NS cells were split as appropriate to maintain a plating density of ~10 cells/mm2; differentiating cells were maintained in media supplemented with bone morphogenic protein (BMP) without EGF and FGF-2. After 8 daysâ treatment cells were harvested, counted and pelleted in aliquots of 50,000 cells. Cells were lysed and the nuclei were pelleted then suspended in buffer and treated with Tn5 transposase for 30 min. ATAC library preparation was undertaken according to published protocols (Buenrostro et al. 2013, Nat Methods 10:1213-8). DNA was purified using Qiagen MinElute columns, followed by 12 cycles of PCR amplification using NEBNext High Fidelity Master Mix and custom primers, with adapter sequences previously specified for ATAC-seq (Buenrostro et al., 2013), followed by further purification on MinElute columns.
ENA-FIRST-PUBLIC
2015-10-01T17:02:43Z
ENA-LAST-UPDATE
2018-03-09T03:06:43Z
External Id
SAMEA3532034
INSDC center name
European Molecular Biology Laboratory EMBL-EBI
INSDC first public
2015-10-01T17:02:43Z
INSDC last update
2018-03-09T03:06:43Z
INSDC status
public
Submitter Id
E-MTAB-3868:CB11130_BMP_A
broker name
ArrayExpress
cell line
CB11130
cell type
Neural stem cell
common name
human
sample name
E-MTAB-3868:CB11130_BMP_A
scientific_name
Homo sapiens
stimulus
bone morphogenic protein
time
8