Subtractive hybridization strategy using paramagnetic oligo(dT) beads and PCR

Melinda Mészáros, David B. Morton

Research output: Contribution to journalArticlepeer-review

11 Scopus citations


Subtractive hybridization has been widely used for the identification of differentially expressed genes. Here we describe a simple, sensitive strategy of subtractive hybridization that involves binding the driver poly(A)+ RNA pool to paramagnetic Dynabeads® Oligo (dT)25. After hybridization with target cDNA, the molecules common to both pools are removed. The subtracted cDNA is then amplified with PCR and used for library screening. Using this method, we have identified four cDNA clones that represent developmentally regulated transcripts in the central nervous system of the tobacco hornworm Manduca sexta. All four transcripts are of low abundance, comprising only 0.001%-0.5% of the poly(A)+ RNA pool.

Original languageEnglish (US)
Pages (from-to)413-419
Number of pages7
Issue number3
StatePublished - Mar 1996
Externally publishedYes

ASJC Scopus subject areas

  • Biotechnology
  • General Biochemistry, Genetics and Molecular Biology


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