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An STR Melt Curve Genotyping Assay for Forensic Analysis Employing an Intercalating Dye Probe FRET *
Author(s) -
Halpern Micah D.,
Ballantyne Jack
Publication year - 2011
Publication title -
journal of forensic sciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.715
H-Index - 96
eISSN - 1556-4029
pISSN - 0022-1198
DOI - 10.1111/j.1556-4029.2010.01549.x
Subject(s) - genotyping , microsatellite , förster resonance energy transfer , capillary electrophoresis , melting curve analysis , genetics , biology , microbiology and biotechnology , str analysis , allele , computational biology , genotype , fluorescence , polymerase chain reaction , gene , physics , quantum mechanics
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.