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Purification of Low-Abundance Messenger RNAs by Polysome Isolation with Polyclonal Antibodies

  • Jan P. Kraus

Abstract

Current interest in the isolation and characterization of normal and mutant genes has focused attention on the need for recombinant DNA molecules complementary to low-abundance mRNAs. The low-abundance class of mRNAs (i.e., each constituting <0.5% of total cellular mRNA) comprises approximately 30% of the mRNA of the eukaryotic cell and codes for a majority of its enzymes. Starting with a total unfractionated mRNA, a cDNA library of at least 200,000 independent clones is required to obtain a complete representation of low-abundance mRNAs.1 A major problem in recovering a cDNA corresponding to a low-abundance mRNA from such a library is the laborious screening required to dotoct the rare cloned low-abundance sequences. A substantial initial enrichment of a given low-abundance mRNA would simplify dramatically the procedures involved in preparing or identifying recombinant DNA molecules containing complementary nucleotide sequences.

Keywords

Sodium Dodecyl Sulfate Magnesium Acetate Vanadyl Sulfate Nascent Polypeptide Chain Bethesda Research Laboratory 
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

© Plenum Press, New York 1985

Authors and Affiliations

  • Jan P. Kraus
    • 1
  1. 1.Department of Human GeneticsYale University School of MedicineNew HavenUSA

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