Development of a Fast, Simple Profiling Method for Sample Screening Using High Resolution Melting (HRM) of STRs

• NCJ Number: 240849
• Author(s): Eric Buel,, Janice A. Nicklas,, Trisha Noreault-Conti,
• Date Published: March 2012
• Length: 11 pages
• Agencies: NIJ-Sponsored
• Award: Improving the efficiency of DNA casework analysis through simple, effective, PCR-based screening methods

Annotation

This article examines a screening assay developed to provide preliminary individualization of crime scene samples thus eliminating expensive, time-consuming short tandem repeat (STR) profiling of nonprobative samples.

Abstract

A screening assay has been developed to provide preliminary individualization of crime scene samples thus eliminating expensive, time-consuming short tandem repeat (STR) profiling of nonprobative samples. High resolution melting performed in a real-time PCR instrument is used to detect the slight melting differences between the length and sequence variations of 22 forensic STRs. Three STRs (vWA, D18S51, THO1) were chosen to develop an assay which was optimized for Mg++ concentration, annealing/extension time/temperature, assay volume, and bovine serum albumin addition. The assay was tested for reproducibility, uniformity for genotype, melting profile consistency, effects of inhibitors, and mixture effects. The assay could be used to determine DNA concentration when a standard curve is run simultaneously. Calculations of costs show that the assay can save significant time and money for a crime with many samples or suspects. Abstract published by arrangement with John Wiley & Sons.