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Selective isolation of highly polymorphic (dC-dA)n.(dG-dT)n microsatellites by stringent hybridization
1Oncogene Division, National Cancer Center Research Institute, Tokyo, Japan.
Genetic Analysis, Techniques and Applications
|February 1, 1993
Summary
Researchers developed a method to isolate long (dC-dA)n.(dG-dT)n microsatellites from plasmid libraries. This technique efficiently identifies polymorphic DNA sequences, crucial for genetic studies.
Area of Science:
- Molecular Biology
- Genetics
Background:
- Microsatellites, or simple sequence repeats (SSRs), are valuable genetic markers.
- Isolating long and polymorphic microsatellites from genomic libraries can be challenging.
Purpose of the Study:
- To establish a hybridization condition for isolating clones containing long (dC-dA)n.(dG-dT)n microsatellites.
- To assess the efficiency and polymorphism of the isolated microsatellites.
Main Methods:
- Construction of a genomic plasmid library.
- Hybridization using a (dC-dA)20 oligonucleotide probe.
- Application of stringent hybridization conditions.
Main Results:
- A specific hybridization condition was defined for microsatellite isolation.
- Over two-thirds of isolated plasmids contained (dC-dA)n.(dG-dT)n repeats longer than (dC-dA)14.
- The identified microsatellites demonstrated high polymorphism.
Conclusions:
- The developed method is effective for isolating long and polymorphic (dC-dA)n.(dG-dT)n microsatellites.
- This technique facilitates the identification of valuable genetic markers for population genetics and other studies.