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


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.

SUBMITTER: Diatchenko L 

PROVIDER: S-EPMC39182 | biostudies-other | 1996 Jun

REPOSITORIES: biostudies-other

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

Diatchenko L L   Lau Y F YF   Campbell A P AP   Chenchik A A   Moqadam F F   Huang B B   Lukyanov S S   Lukyanov K K   Gurskaya N N   Sverdlov E D ED   Siebert P D PD  

Proceedings of the National Academy of Sciences of the United States of America 19960601 12


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  ...[more]

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