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  • Title: Short tandem repeat typing by capillary array electrophoresis: comparison of sizing accuracy and precision using different buffer systems.
    Author: Vainer M, Enad S, Dolnik V, Xu D, Bashkin J, Marsh M, Tu O, Harris DW, Barker DL, Mansfield ES.
    Journal: Genomics; 1997 Apr 01; 41(1):1-9. PubMed ID: 9126475.
    Abstract:
    Polymorphic microsatellite markers are widely used in gene discovery and mapping, human identification, agricultural genetics, and diagnosis of triplet-repeat expansion disorders. Reliable genotyping of these markers requires polymerase chain reaction (PCR) amplification and very-high-resolution electrophoresis. Capillary array electrophoresis offers extremely fast, high-resolution separation of DNA and more automated sample processing because labor-intensive slab-gel pouring and sample loading are eliminated. We report a simple, reliable procedure for preparing PCR samples for electrokinetic injection into capillaries using a 96-well tray and float dialysis. We developed an improved sizing standard for genotyping and used it to evaluate systematically the sizing accuracy and precision of low-viscosity, replaceable matrix formulations. Our study sizing over 28,000 alleles yielded an average precision of +/- 0.12 bp for fragments up to 350 bp. Low-viscosity formulations permit low-pressure matrix injection (40 psi) and a turnaround time of 70 min for 48-96 samples.
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