DNA from cell lines was purchased from the Coriell Institute for Medical Research (Camden, NJ).
Label
Biotin
Label protocol
Standard Affymetrix protocol.
Hybridization protocol
The DNA samples are hybridized to the arrays at 49oC in a buffer that contains MES (58 mM), Denhardt's solution (2.5X), EDTA (6 mM), Herring Sperm DNA (0.12 mg/ml), Human Cot-1 DNA (0.01 mg/ml), Tween-20 (0.01%), DMSO (5 %), and TMACl (2.7M). The arrays are then washed using Wash A (6X SSPE, 0.01% Tween 20) at 25oC followed by washing with Wash B (0.6X SSPE, 0.01% Tween 20)at 45oC. The arrays are then stained with streptavidin conjugated to phycoerythrin (SAPE) for 10 minutes at 25oC, followed by staining with a biotinylated goat Anti-streptavidin antibody for 10 minutes at 25oC, followed by another stain with SAPE at 25oC. There is then a final wash with Wash A at 30oC.
Scan protocol
The arrays are scanned on a GCS3000 scanner (Affymetrix).
Description
Hapmap sample NA10846 was hybridized to Nsp CN arrays
