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Nucleic Acids Research, Vol 26, Issue 15 3616-3618, Copyright © 1998 by Oxford University Press

ARTICLES

Discovery of estrogen-responsive genes using an improved method which combines subtractive hybridization and PCR

W Liu, W Su and TM Roberts
Division of Cellular and Molecular Biology, Dana-Farber Cancer Institute and Department of Pathology,Harvard Medical School, Boston, MA 02115, USA.

Here we describe a reliable method for isolating genes that are differentially expressed in two cell populations. The method is a combination of subtractive hybridization and PCR. Among many improvements to previously described methods is the incorporation of a new technology into the procedure which sterilizes(inactivates) PCR amplicons, and thereby overcomes the limitation of similar procedures. To test this improved method, we conducted a search for estrogen- responsive genes. Estrogen-regulated genes dominated the subtracted libraries after four rounds of subtractive hybridizations. Four estrogen-regulated genes were identified from the initial screening.
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