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. 1996 Jun 11;93(12):6025-30.
doi: 10.1073/pnas.93.12.6025.

Suppression Subtractive Hybridization: A Method for Generating Differentially Regulated or Tissue-Specific cDNA Probes and Libraries

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Free PMC article

Suppression Subtractive Hybridization: A Method for Generating Differentially Regulated or Tissue-Specific cDNA Probes and Libraries

L Diatchenko et al. Proc Natl Acad Sci U S A. .
Free PMC article

Abstract

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.

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References

    1. Science. 1983 Oct 14;222(4620):135-9 - PubMed
    1. Nucleic Acids Res. 1995 Nov 25;23(22):4738-9 - PubMed
    1. Proc Natl Acad Sci U S A. 1984 Apr;81(7):2194-8 - PubMed
    1. Proc Natl Acad Sci U S A. 1988 Aug;85(15):5738-42 - PubMed
    1. Cytometry. 1990;11(1):208-18 - PubMed

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