Simple and efficient subtractive hybridization screening

Biotechniques. 1991 Nov;11(5):598, 600, 602-4.

Abstract

A method of subtractive hybridization screening is presented that does not rely on the availability of large amounts of poly(A)+ RNA. The usefulness of this method is demonstrated by the isolation of cDNA clones that are unique or present at enhanced levels in cDNA libraries derived from a psoriatic skin cell line. A very high level of enrichment for rare cDNA sequences is possible as shown by the isolation of cDNA clones present at a frequency of less than one in one hundred thousand (0.001%). This method is particularly useful in experimental systems where the amount of RNA available is limited.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Blotting, Northern
  • Blotting, Southern
  • Cloning, Molecular
  • DNA / isolation & purification*
  • Humans
  • Molecular Sequence Data
  • Nucleic Acid Hybridization*
  • Oligonucleotide Probes
  • Polymorphism, Restriction Fragment Length

Substances

  • Oligonucleotide Probes
  • DNA