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A simple method for filter hybridization of labeled restriction fragments excised from gels
Authors:R Marotta  M Bremenkamp  F Salamini
Institution:Istituto Sperimentale per la Cerealicoltura, Sezione di Bergamo, via Stezzano 24, P.O. Box 164, 24100 Bergamo, Italy
Abstract:Labeled DNA restriction fragments excised from agarose or bisacrylylcystamine-acrylamide gels can be used for hybridization to nitrocellulose-bound DNA without eliminating the gel matrix. A gel slice containing the labeled fragment is excised, dissolved by heating at 105 degrees C (in the presence of beta-mercaptoethanol for bisacrylylcystamine-acrylamide gels), and added to the hybridization mixture. The presence of agarose or polyacrylamide in the solution does not inhibit hybridization. The method is simple, rapid, and allows complete recovery of the probe.
Keywords:agarose  bisacrylylcystamine-acrylamide  restriction fragment  end labeling  filter hybridization  pBR 322
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