4.5 Article

Interrogation of short tandem repeats using fluorescent probes and melting curve analysis: A step towards rapid DNA identity screening

Journal

FORENSIC SCIENCE INTERNATIONAL-GENETICS
Volume 2, Issue 4, Pages 333-339

Publisher

ELSEVIER IRELAND LTD
DOI: 10.1016/j.fsigen.2008.04.007

Keywords

Short tandem repeats (STRs); Fluorescent probe; HyBeacon; Melting curve analysis; Real-time PCR

Funding

  1. Engineering and Physical Sciences Research Council [EP/D036453/1] Funding Source: researchfish

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Current forensic DNA profiling methods rely on the analysis of samples at specialised laboratories with an average turnaround time of several days. The ability to rapidly determine a partial profile of short tandem repeats at the point-of-arrest would be of great benefit to police forces around the world, for example enabling a suspect to be rapidly included or excluded from an investigation. We have developed a homogeneous PCR method for the interrogation of STR loci utilising fluorescent oligonucleotide probes and melting Curve analysis. Alleles of the D18S51, TH01 and D8S1179 loci were differentiated and identified oil the basis of tat-get length and probe melting temperature. Assay performance was evaluated by comparing melting peak data with the AmpFISTR(R) SGM Plus(R) system. The method is compatible with direct analysis of unpurified buccal swab samples, enabling a partial STR profile to be generated within 1 h. (C) 2008 Elsevier Ireland Ltd. All rights reserved.

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