7 protocols
AccessionNameType
P-MTAB-28183
image_acquisition
Scanning was carried out according to manufacturer specifications.
P-MTAB-28182
hybridization
Labeled genomic DNA fragments were hybridized to Affymetrix Genome-Wide Human SNP 6.0 arrays.
P-MTAB-28181
Labeling
500 ng of genomic DNA was subjected ty restriction enzyme digestion by NspI and StyI and ligated to adapters. Ligation products were PCR amplified with a biotinylated primer, fragmented and labeled with Cy3 for single-channel hybridization.
P-MTAB-28178
grow
iPS cells were cultured in DMEM/F-12 supplemented with: non-essential amino acids, 2mM L-Glutamine, 0.1mM 2-Mercaptoethanol, 20% Knockout Serum Replacement (Life Technologies) and FGF-2 (10ng/ml). Cells were plated on mitotically-inactivated mouse embryonic fibroblasts. Cells were split every 7-14 days (1:3-1:10) or frozen in clumps using dissociation buffer (20% KSR, 1mM CaCl2, 1mg/ml Collagenase IV, 0.25% Trypsin in PBS) or freezing buffer (2M DMSO, 1M Acetamide, 3M propylene glycol in ES cell media).
P-MTAB-28179
specified_biomaterial_action
Cells were dissociated using Accumax (Millipore) and feeder cells depleted.
P-MTAB-28180
nucleic_acid_extraction
Genomic DNA was extracted using the DNA Wizard kit (Promega).
P-MTAB-28177
grow
GNS cells were cultured using serum-free basal medium supplemented with B27 and N2 (Life Technologies) and growth factors EGF and FGF-2 (20 ng/ml). Culture vessels were coated with Laminin (Sigma) for 3 h 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.