Human embryonic stem cells derived by Cellartis AB grown on STO feeders. Manual dissection was systematically used to passage the cells rather than enzymatic methods.
Extracted molecule
genomic DNA
Extraction protocol
Genomic DNA was isolated from hESC using the Wizard® Genomic DNA Purification Kit (Promega, Charbonnieres, France).
Label
CY3
Label protocol
A genomic DNA labeling kit (Enzo, Farmingdale, NY) was used to label the sample DNA with Cy5. The labeled products were purified with QiaQuick polymerase chain reaction (PCR) purification kits (Qiagen, Valencia, CA).
Research Centre for Reproduction and Genetics, University Hospital and Medical School of the Vrije Universiteit Brussel, Brussels, Belgium.
Growth protocol
Human embryonic stem cells derived as described by Mateizel, I. et al. Hum. Reprod. 21, 503-511 (2006) and grown on STO feeders. Manual dissection was systematically used to passage the cells rather than enzymatic methods.
Extracted molecule
genomic DNA
Extraction protocol
Genomic DNA was isolated from hESC using the Wizard® Genomic DNA Purification Kit (Promega, Charbonnieres, France).
Label
CY5
Label protocol
A genomic DNA labeling kit (Enzo, Farmingdale, NY) was used to label the sample DNA with Cy3. The labeled products were purified with QiaQuick polymerase chain reaction (PCR) purification kits (Qiagen, Valencia, CA).
Hybridization protocol
After addition of tRNA (48 mg) plus a quantity of human Cot DNA (Invitrogen, Carlsbad, CA) equal to 50 times the mass of the labeled DNAs, the resulting products were concentrated by vacuum centrifugation, resuspended in 43 mL of hybridization solution (Ambion, Austin, TX) and deposited on the array under a coverslip. Hybridization in a sealed chamber (Corning Life Sciences, Acton, MA) was for 66 hours at 55_C, a post hybridization treatement was done.
Scan protocol
Slides were scanned (Agilent Technologies, Santa Clara, CA), data were extracted with GenePix 6 (Axon InstrumentseMolecular Devices, Sunnyvale, CA),
Description
DATA exportation with Genocensus software
Data processing
All data is lowess normalized, ratio value is CY5/CY3. Copy number analysis was performed with GenoCensus (IntegraGen), which includes block Loess normalization
