Title

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

Authors

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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