Suppression subtractive hybridization: a method for generating differentially regulated or tissue-specific cDNA probes and libraries.
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.
Duke Scholars
Altmetric Attention Stats
Dimensions Citation Stats
Published In
DOI
ISSN
Publication Date
Volume
Issue
Start / End Page
Location
Related Subject Headings
- Y Chromosome
- Nucleic Acid Hybridization
- Molecular Sequence Data
- Humans
- Genomic Library
- Gene Expression Regulation
- DNA, Complementary
- DNA Probes
- Cosmids
- Chromosome Mapping
Citation
Published In
DOI
ISSN
Publication Date
Volume
Issue
Start / End Page
Location
Related Subject Headings
- Y Chromosome
- Nucleic Acid Hybridization
- Molecular Sequence Data
- Humans
- Genomic Library
- Gene Expression Regulation
- DNA, Complementary
- DNA Probes
- Cosmids
- Chromosome Mapping