Application of automated DNA sizing technology for genotyping microsatellite loci
- Applied Biosystems, Foster City, CA (United States)
- Virginia Commonwealth Univ., Richmond, VA (United States)
Highly polymorphic microsatellite loci offer great promise for gene mapping studies, but fulfillment of this potential will require substantial improvements in methods for accurate and efficient genotyping. Here, the authors report a genotyping method based on fluorescently labeled PCR primers and size characterization of PCR products using an automated DNA fragment analyzer. They capitalize on the availability of three distinct fluorescent dyes to uniquely label loci that overlap in size. This innovation increases by threefold the number of loci that can be analyzed simultaneously. They label size standards with a fourth dye and combine these with the microsatellite PCR products in each gel lane. Computer programs provide very rapid and accurate sizing of microsatellite alleles and efficient data management. In addition, fluorescence signals are linear over a much greater range of intensity than conventional autoradiography. This facilitates multiplexing of loci (since signal intensities often vary greatly) and helps distinguish major peaks from artifacts, thereby improving genotyping accuracy. 29 refs., 3 figs.
- OSTI ID:
- 6758292
- Journal Information:
- Genomics; (United States), Journal Name: Genomics; (United States) Vol. 14:4; ISSN GNMCEP; ISSN 0888-7543
- Country of Publication:
- United States
- Language:
- English
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