Suppression subtractive hybridization: a method for generating differentially regulated or tissue-specific cDNA probes and libraries.

Journal Article (Journal Article)

A new and highly effective method, termed suppression subtractive hybridization (SSH), has been developed for the generation of subtracted cDNA libraries. It is based primarily on a recently described technique called suppression PCR and combines normalization and subtraction in a single procedure. The normalization step equalizes the abundance of cDNAs within the target population and the subtraction step excludes the common sequences between the target and driver populations. In a model system, the SSH technique enriched for rare sequences over 1,000-fold in one round of subtractive hybridization. We demonstrate its usefulness by generating a testis-specific cDNA library and by using the subtracted cDNA mixture as a hybridization probe to identify homologous sequences in a human Y chromosome cosmid library. The human DNA inserts in the isolated cosmids were further confirmed to be expressed in a testis-specific manner. These results suggest that the SSH technique is applicable to many molecular genetic and positional cloning studies for the identification of disease, developmental, tissue-specific, or other differentially expressed genes.

Full Text

Duke Authors

Cited Authors

  • Diatchenko, L; Lau, YF; Campbell, AP; Chenchik, A; Moqadam, F; Huang, B; Lukyanov, S; Lukyanov, K; Gurskaya, N; Sverdlov, ED; Siebert, PD

Published Date

  • June 11, 1996

Published In

Volume / Issue

  • 93 / 12

Start / End Page

  • 6025 - 6030

PubMed ID

  • 8650213

Pubmed Central ID

  • PMC39182

International Standard Serial Number (ISSN)

  • 0027-8424

Digital Object Identifier (DOI)

  • 10.1073/pnas.93.12.6025


  • eng

Conference Location

  • United States