7 protocols
AccessionType
bioassay_data_transformation
ID_REF = VALUE = MAS5.0 signal ABS_CALL = DETECTION P-VALUE =
feature_extraction
Microarray Analysis Suite 5.0 (MAS, Affymetrix) was used to quantify microarray signals with default analysis parameters and global scaling to target MEAN = 150
nucleic_acid_extraction
Total RNA was isolated uisng the PicoPure RNA isolation kit (Arcturus). Isolated RNA was subsequently amplified using GeneChip® Eukaryotic Small Sample Target Labeling Assay Version II
image_aquisition
Default Affymetrix GCOS settings for GeneChip 3000 7G scanner
grow
All mouse embryos were collected and washed in M2 medium and incubated in modified KSOM under mineral oil in a humidfied atmosphere of 5% CO2, 5% O2, 90% N2 atmosphere at 37°C until use
labeling
Biotinylated cRNA were prepared using ENZO BioArray HighYied RNA transcript labeling Kit according to the manufacturer's instructions.
hybridization
aRNA was hybridized for 16 hr on Affymetrix mouse 430 v2.0 microarrays. GeneChips were washed and stained using an Affymetrix Fluidics Station 450.