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STR Melting Curve Analysis as a Genetic Screening Tool for Crime Scene Samples *
Author(s) -
Nguyen Quang,
McKinney Jason,
Johnson Donald J.,
Roberts Katherine A.,
Hardy Winters R.
Publication year - 2012
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.1556-4029.2012.02106.x
Subject(s) - amplicon , melting curve analysis , genotyping , genotype , dna profiling , dna , standard curve , microsatellite , biology , microbiology and biotechnology , chromatography , computational biology , analytical chemistry (journal) , polymerase chain reaction , genetics , chemistry , gene , allele
In this proof‐of‐concept study, high‐resolution melt curve (HRMC) analysis was investigated as a postquantification screening tool to discriminate human CSF1PO and THO1 genotypes amplified with mini‐STR primers in the presence of SYBR Green or LCGreen Plus dyes. A total of 12 CSF1PO and 11 HUMTHO1 genotypes were analyzed on the LightScanner HR96 and LS‐32 systems and were correctly differentiated based upon their respective melt profiles. Short STR amplicon melt curves were affected by repeat number, and single‐source and mixed DNA samples were additionally differentiated by the formation of heteroduplexes. Melting curves were shown to be unique and reproducible from DNA quantities ranging from 20 to 0.4 ng and distinguished identical from nonidentical genotypes from DNA derived from different biological fluids and compromised samples. Thus, a method is described which can assess both the quantity and the possible probative value of samples without full genotyping.