STR Melting Curve Analysis as a Genetic Screening Tool for Crime Scene Samples


  • Presented in part at the Promega 17th International Symposium on Human Identification, October 10, 2006, in Nashville, TN.

  • Present address: VA Medical Center Center for Regenerative Medicine, C-3113, 1481 West 10th Street, Indianapolis, IN 46202.

Additional information and reprint requests:
Winters Reef Hardy, Ph.D.
VA Medical Center
Center for Regenerative Medicine, C-3113
1481 West 10th Street
Indianapolis, IN 46202


Abstract:  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.