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

NCJ Number
Journal of Forensic Sciences Volume: 56 Issue: 1 Dated: January 2011 Pages: 36-45
Date Published
January 2011
10 pages

This study focused on an advancement within the category of hybridization-based detection.


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 less than 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. (Published Abstract) Table, figures, and references

Date Published: January 1, 2011