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- W217877204 abstract "Short tandem repeat polymorphisms (STRPs) including di-, tri-, and tetranucleotide repeats are very useful markers for gene mapping, genetic diagnosis, and forensics because they are highly polymorphic, abundant throughout the mammalian genome, and amplifiable by the polymerase chain reaction (PCR). In order to meet the demand for large scale gene mapping and genetic diagnosis, the overall speed of genotyping STRPs should be increased. Automated detection systems using laser irradiation along with infrared fluorescently labeled PCR primers or dATP`s have been used for improved detection of PCR amplified STRPs when compared to conventional radioactive detection methods. Here, we report protocols that provide high throughput visualization and analysis of PCR amplified STRPs using the LI-COR Model 4000S DNA Sequencer. Short (15 cm separation distance) gels were used which produced rapid migration of the DNA fragments to the scanning detector (less than 1 hour from sample loading to detection of up to 350 base long DNA fragments). Seven percent denaturing acrylamide gels with a constant 2000V were employed for fast and adequate resolution. A 64 well format was used for loading (60 samples with 4 lanes for standard markers). Multiple loading of samples (using the same gel up to 3 times) has beenmore » achieved. In addition, we have multiplexed more than one locus per lane. By applying these conditions (60 samples x 3 loci x loads/gel x 2 gels/day) it is possible to generate images for over 1000 person loci in less than a day. Images were analyzed using Scanalytics RFLPscan software. The output data gave each allele size in number of base pairs.« less" @default.
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- W217877204 date "1994-09-01" @default.
- W217877204 modified "2023-09-27" @default.
- W217877204 title "High throughput visualization and analysis of human short tandem repeat polymorphisms (STRP`s) using an infrared based automated DNA sequencer" @default.
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