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Applied Biosystems Applied Biosystems fluorescent multicolor dye technology allows multiple loci,
Fluorescent Dye including loci that have alleles with overlapping size ranges, to be analyzed in a single
Technology gel lane or capillary injection. Alleles for overlapping loci are distinguished by labeling
locus-specific primers with different color dyes.
Because only one primer of each pair is labeled, the ABI PRISM® instruments detect
only one strand for each amplified DNA fragment. The detection of only one strand
eliminates doublets arising from the different mobilities of complementary strands that
are often observed when using gel staining detection methods.
Automated Sizing Amplified samples can be analyzed in a slab gel format or can be injected into a
and Genotyping capillary. An internal lane size standard is loaded with each sample to allow for
automatic sizing of the PCR products and to normalize differences in electrophoretic
mobility between gel lanes or injections (Ziegle et al., 1992). GeneScan software
automatically analyzes the collected data, which can then be imported into Genotyper
software for automatic genotyping of alleles.
High Throughput Laboratories can analyze hundreds of loci in a single day using four-dye fluorescent
labeling. This is a dramatic increase in productivity compared with gel staining
techniques, which visualize all PCR products in the same color, or with other systems
that are limited to one or two colors.
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9-20 Microsatellite Analysis Applications