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Monitoring Gene Expression Using DNA Arrays
Abstract
A single phenotypic change observed during an organism’s growth or adaptation to its environment is usually the result of a coordinated expression of genes ranging from a few to many. Various techniques are in use today for quantifying gene expression. These include indirect methods, such as protein reporter fusions (e.g., β-galactosidase, chloramphenicol acetyltransferase, green fluorescent protein, and luciferase), and more direct techniques such as Northern blotting. Nevertheless, these techniques suffer from the drawbacks of being time-consuming, labor-intensive and allow only small numbers of genes to be studied at any given moment. DNA array technology now enables the analysis of a larger number of genes, including whole genomes, in a single experiment (1). In general, DNA samples of genes of interest are spotted onto a solid support surface in an ordered manner. RNA is isolated from both the control and experimental bacterial samples and reverse-transcribed while simultaneously incorporating marker molecules (radioactive or Cy3 and/or Cy5 nucleotides), resulting in labeled cDNA. The degree of hybridization of the labeled cDNA probes to the arrays is then detected using appropriate systems. A comparison is made of the intensities of the probe between the control and experimental samples and any differences observed reflect either up- or downregulation of particular genes.
Affiliation(s): (2) Department of Pediatrics, Faculty of Medicine, Imperial College of Science, Technology and Medicine, London, UK
(3) Department of Pediatrics, Faculty of Medicine, Imperial College of Science, Technology and Medicine, St. Mary’s Hospital, London, UK
Series: Methods in Molecular Medicine  |  Volume: 71  |  Pub. Date: Sep-25-2002  |  Page Range: 119-134  |  DOI: 10.1385/1-59259-321-6:119
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