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PCR Cloning Protocols pp 389–403Cite as

Generation of a PCR-Renewable Source of Subtractive cDNA

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Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 67))

Abstract

Differential (+/-) first-strand cDNA screening methods identify clones corresponding to mRNAs that are expressed at a higher level in one of a pair of phenotypically different cells. This approach is limited by the fact that screening of libraries with labeled first-strand cDNAs synthesized from unfractionated mRNA can detect clones containing sequences representing approx 0.1% or more of the complexity of mRNA (i.e., mRNAs present at greater than about 200 copies/cell since a typical mammalian cell line contains approx 250,000 mRNAs).

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Author information

Authors and Affiliations

  1. NCI-Navy Medical Oncology, Bethesda, MD

    W. Michael Kuehl

  2. NCI-Naval Medical Oncology, Bethesda, MD

    James Battey

Authors
  1. W. Michael Kuehl
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  2. James Battey
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Editor information

Editors and Affiliations

  1. Department of Anatomy, University of Connecticut Health Center, Farmington, CT

    Bruce A. White

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© 1997 Humana Press Inc.

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Kuehl, W.M., Battey, J. (1997). Generation of a PCR-Renewable Source of Subtractive cDNA. In: White, B.A. (eds) PCR Cloning Protocols. Methods in Molecular Biology™, vol 67. Humana Press, Totowa, NJ. https://doi.org/10.1385/0-89603-483-6:389

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  • DOI: https://doi.org/10.1385/0-89603-483-6:389

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-0-89603-483-9

  • Online ISBN: 978-1-59259-553-2

  • eBook Packages: Springer Protocols

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