Strain: C57BL/6J Gender: Male Age: 10 weeks Tissue: intestinal mucosa
Extracted molecule
total RNA
Label
R-phycoerythrin
Description
Eight micrograms of total RNA were converted into double-stranded cDNA (ds-cDNA) by use of the SuperScript double-stranded cDNA synthesis kit and GeneChip T7-oligo(dT) promoter primer kit. After second-strand synthesis, the reaction mixture was purified using the GeneChip sample cleanup module. In vitro transcription was performed on the ds-cDNA with the Enzo RNA transcript labeling kit. Biotinylated cRNA was purified using the GeneChip sample cleanup module and fragmented by incubating 94°C for 35 min. The biotinylated cRNA (final 0.05 mg/ ml) was hybridized with Mouse Expression Array 430A for 16 h at 45°C. After stringent washing in a microfluidics station 400, cRNA was stained with R-phycoerythrin streptavidine and scanned after antibody amplification. Fluorescence intensities were measured with an Affymetrix Scanner. Image output files were inspected for hybridization artifacts, and based on the fluorescence intensity differences between perfect match and mismatch probes, a hybridization intensity was calculated for each transcript using Microarray Suite 5.0. Default parameters optimized by the manufacturer for this microarray chip were used for all signal analysis.
Data processing
Normalized by the value of acidic ribosomal phosphoprotein PO (1419441_at)
Differential regulation of intestinal lipid metabolism in A/J versus C57BL/6J mice during diet-induced obesity
Data table header descriptions
ID_REF
VALUE
Signal value that represents relative measure of transcript abundance
ABS_CALL
The call in an absolute analysis that indicates if the transcript was present (P), absent (A), or marginal (M)
LOG_RATIO_B6LF
Signal log ratio in a comparison analysis on the signal for C57BL/6J mice fed a low fat diet (GSM77382)
CHANGE_CALL_B6LF
The call in a comparison analysis on the signal for C57BL/6J mice fed a low fat diet (GSM77382), which indicates if the signal log ratio was increased (I), marginally increased (MI), decreased (D), marginally decreased (MD), or not changed (NC)
