An STR Melt Curve Genotyping Assay for Forensic Analysis Employing an Intercalating Dye Probe FRET

    Authors

    M. D. Halpern;J. Ballantyne

    Comments

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    Abbreviated Journal Title

    J. Forensic Sci.

    Keywords

    forensic science; DNA typing; short tandem repeat; fluorescence; resonance energy transfer; intercalating dye; fluorophore probe; melt; curve analysis; RESONANCE ENERGY-TRANSFER; DNA; HYBRIDIZATION; MICROARRAYS; LOCI; PCR; Medicine, Legal

    Abstract

    The most common markers used in forensic genetics are short tandem repeats (STRs), the alleles of which are separated and analyzed by length using capillary electrophoresis (CE). In this work, proof of concept of a unique STR genotyping approach has been demonstrated using asymmetric PCR and a fluorescence resonance energy transfer (FRET)-based hybridization analysis that combines fluorophore-labeled allele-specific probes and a DNA intercalating dye (dpFRET) in a melt match/mismatch analysis format. The system was successfully tested against both a simple (TPOX) and a complex (D3S1358) loci, demonstrated a preliminary detection limit of < 10 genomic equivalents with no allelic dropout and mixture identification in both laboratory-generated and clinical samples. With additional development, this approach has the potential to contribute to advancing the use of STR loci for forensic applications and related fields.

    Journal Title

    Journal of Forensic Sciences

    Volume

    56

    Issue/Number

    1

    Publication Date

    1-1-2011

    Document Type

    Article

    Language

    English

    First Page

    36

    Last Page

    45

    WOS Identifier

    WOS:000285839300006

    ISSN

    0022-1198

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