Discovery of estrogen-responsive genes using an improved method which combines subtractive hybridization and PCR.
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.
Division of Cellular and Molecular Biology, Dana-Farber Cancer Institute and Department of Pathology,Harvard Medical School, Boston, MA 02115, USA.
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Roberts
D004967:Estrogens / Q000378:metabolism* / Q000494:pharmacology; D006801:Humans; D009693:Nucleic Acid Hybridization / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D014407:Tumor Cells, Cultured
D004967:Estrogens