tissue: cardiac age: 3 month old group: EX state: at rest
Extracted molecule
total RNA
Extraction protocol
Global gene expression was measured using RNA from the frozen heart tissue. Briefly, RNA was extracted from 20-30 mg samples cut from the left ventricles using Qiagen RNeasy (Valencia, CA) RNA extraction kits. The absorbance ratio (A260/A280) was determined using a ND-1000 UV/VIS spectrophotometer (Nanodrop Technologies, Inc., Montchanin DE) to verify that the RNA was of high purity. The integrity of each sample was further assessed with an Agilent BioAnalyzer system by the Biomedical Genomics Core at the Research Institute at Nationwide Childrens Hospital (Columbus, OH). Samples passing quality control were pooled within each treatment group to yield 4 gene chips per treatment group, with RNA from 3 animals on each chip, to ensure correct quantity of total RNA.
Label
biotin
Label protocol
The pooling of RNA samples allowed for representation in a cost-effective analyses. Samples were labeled with the Affymetrix® Whole Transcript Labeling system
Hybridization protocol
hybridized to the Affymetrix GeneChip® Rat Gene 1.0 ST Array (Santa Clara, CA).
Scan protocol
Affymetrix scan protocol
Description
Cardiac gene expression data from 3 month old EX group at rest
Data processing
The design of the Rat Gene 1.0 ST Array is based primarily on a subset of GeneChip® Rat Exon 1.0 ST Array probes that map to well-supported exons of known genes. The array comprised of more than 700,000 unique 25-mer oligonucleotide features constituting more than 27,000 gene-level probe sets. Data was preprocessed using the RMA approach for background correction, normalization and probe set summarization using the Bioconductor affy package in R.
Gene expression data from 16-month and 3-month old male rats, with three different levels of activity over the life span: sedentary, twice weekly physical activity, regular running wheel exercise.
