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Screening cDNA Libraries by Hybridization with Double-Stranded DNA Probes and Oligonucleotides

  • Caroline A. Austin
Protocol
  • 230 Downloads
Part of the Springer Protocols Handbooks book series (SPH)

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, e.g., degenerate probes may be obtained by back translating a peptide sequence of an unknown protein, or they 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. This chapter concentrates on the methods for labeling DNA probes and hybridization to filter-bound library DNA.

Keywords

Sodium Dodecyl Sulfate Standard Saline Citrate Plasmid Library Label Oligonucleotide Probe Prehybridization Buffer 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Humana Press Inc., Totowa, NJ 2000

Authors and Affiliations

  • Caroline A. Austin
    • 1
  1. 1.Department of Biochemistry and Genetics, The Medical SchoolThe University of Newcastle upon TyneNewcastleUK

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