Monitoring eukaryotic gene expression using oligonucleotide microarrays.
An increasing number of biological and medical research questions depend on obtaining global views of gene expression. In this chapter, we will describe how oligonucleotide microarrays have been used to accomplish this goal. In particular, we will focus on the use of GeneChip arrays, which provide high levels of reproducibility, sensitivity, and specificity. Target preparation, hybridization, washing, signal detection, and data analysis will be described in detail. Additionally, we will discuss options for facilitating data sharing, including the creation of databases, and the use of internet tools that help users place their results in the context of data from public and proprietary databases. There is so much interest and innovation in the field of genomics that protocols are constantly evolving. This chapter should be used as a genomic profiling guide only. We urge readers to consult www.affymetrix.com for the most current products and protocols.