Affymetrix Oligonucleotide Analysis of Gene Expression in the Injured Heart

Microarrays have helped researchers gain much insight into gene expres sion profiles in the context of many diseases including those in the injured heart. Our genomic investigations have been focused on elucidation of host gene responses to enterovirus infection. We have gained valuable technical expertise in using Affymetrix oligonucleotide arrays, also known as GeneChipEs, and cDNA spotted arrays to probe differential gene expression in both cultured cells and in heart tissue. Here, we provide a technique-focused supplement to the Affymetrix GeneChip� Expression Analysis Manual for sample preparation, processing, and array hybridization. We provide expanded explanations to highlight important points within the existing protocol and of fer variations to standard procedures when appropriate. For investigators using myocardial tissues for microarray experiments, we further address the neces sity of and methods for in situ flushing of the vasculature, tissue homogeniza tion, and considerations for limits of expression detection in rare cells. It is our intention to provide useful technical information, based on our experience, to assist those researchers using Affymetrix GeneChips in their own genomic re search.