PMID: 9445489Feb 21, 1998Paper

High-precision genotyping by denaturing capillary electrophoresis

Genome Research
H M WenzJ W Efcavitch

Abstract

Genotyping, as applied to linkage mapping, human identification, or mapping of genetic traits, mandates electrophoretic separation systems that enable a user to identify alleles with high precision to obtain a correct genotype. For 2-bp microsatellites or short tandem repeats (STRs), standard deviations of +/-0.3 nucleotide are required to ensure with 99.7% probability the identity or dissimilarity of tested alleles. A complete system, consisting of commercially available laser-induced fluorescence capillary electrophoresis (ABI PRISM 310) and performance optimized polymer 4 (POP-4), was evaluated for microsatellite separations. POP-4 is a low viscosity polymer for use in uncoated fused microbore silica capillaries. It separates DNA fragments that differ in size by 1 nucleotide up to 250 nucleotides and that differ in size by 2 nucleotides for fragments up to at least 350 nucleotides in length in about 30 min. The presence of denaturants and, more importantly, operation at 60 degrees C was mandatory for high-precision and high-resolution sizing operation. Reproducible separation performance was achieved in excess of 100 injections per capillary with resulting standard deviations in the range of 0.04 to 0.17 nucleotide. Comparat...Continue Reading

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Citations

Sep 24, 2004·Proceedings of the National Academy of Sciences of the United States of America·Jonathan R HartJacqueline K Barton
Oct 29, 2009·Electrophoresis·Stefanie SchindlerKarol Szafranski
May 28, 2010·The Journal of Immunology : Official Journal of the American Association of Immunologists·Alan GillettMaja Jagodic
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