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Screening cDNA Libraries by Hybridization with Double-Stranded DNA Probes and Oligonucleotides
Abstract
Probably the most commonly used method to screen a cDNA library is hybridization to a labeled DNA probe. This probe may be a single-stranded oligonucleotide or a double-stranded cDNA or polymerase chain reaction (PCR) product. The DNA may be either radioactively or nonradioactively labeled. The sequence of an oligonucleotide probe may be derived from a number of sources, for example, degenerate probes may be obtained by back-translating a peptide sequence of an unknown protein, or may be a short conserved region of sequence within a cDNA from another member of a multigene family or from a cognate cDNA from another species (see Note 1). Double-stranded DNA probes may be a partial cDNA obtained by screening another library, or a PCR product, or a cDNA from another member of a gene family or from another species.
Affiliation(s): (2) Department of Biochemistry and Genetics, The Medical School University of Newcastle, Newcastle Upon Tyne, UK
Series: Methods in Molecular Biology  |  Volume: 69  |  Pub. Date: Oct-01-1996  |  Page Range: 147-154  |  DOI: 10.1385/0-89603-383-X:147
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