Preparation of a differentially expressed, full-length cDNA expression library by RecA-mediated triple-strand formation with subtractively enriched cDNA fragments

Nucleic Acids Res. 1996 Sep 1;24(17):3478-80. doi: 10.1093/nar/24.17.3478.

Abstract

We have developed a fast and general method to obtain an enriched, full-length cDNA expression library with subtractively enriched cDNA fragments. The procedure relies on RecA-mediated triple-helix formation of single-stranded cDNA fragments with a double-stranded cDNA plasmid library. The complexes were then captured from the solutions using the digoxigenin-antidigoxigenin paramagnetic beads followed by recovery of the enriched double-stranded cDNA expression library. We have observed a linear relation between the capture of full-length cDNAs in the library and the fold enrichment in the subtracted cDNA population.

MeSH terms

  • Affinity Labels
  • Animals
  • Cloning, Molecular / methods*
  • DNA
  • DNA, Complementary / genetics*
  • Digoxigenin
  • Gene Expression*
  • Gene Library*
  • Liver Regeneration
  • Nucleic Acid Conformation
  • Nucleic Acid Hybridization
  • Rats
  • Rec A Recombinases

Substances

  • Affinity Labels
  • DNA, Complementary
  • triplex DNA
  • DNA
  • Rec A Recombinases
  • Digoxigenin