ISSN: 0022-1198
CODEN: JFSCA
Published Online: 5 May 2004
Page Count: 8

A Study on the Effects of Degradation and Template Concentration on the Amplification Efficiency of the STR Miniplex Primer Sets
McCord, BR
Ohio University, OH

Chung, DT
Ohio University, OH

Butler, JM
National Institute of Standards and Technology, Biotechnology Division, MD

Drábek, J
Ohio University, OH

Opel, KL
Ohio University, OH

(Received 10 August 2003; accepted 15 March 2004)

Abstract

In forensic DNA analysis, the samples recovered from the crime scene are often highly degraded leading to poor PCR amplification of the larger sized STR loci. To avoid this problem, we have developed STR markers with redesigned primer sequences called "Miniplexes" to produce smaller amplicons. To assess the effectiveness of these kits, we have tested these primer sets with enzymatically degraded DNA and compared the amplifications to a commercial kit. We also conducted sensitivity and peak balance studies of three Miniplex sets. Lastly, we report a case study on two human skeletal remain samples collected from different environmental conditions. In both types of degraded DNA, the Miniplex primer sets were capable of producing more complete profiles when compared to the larger sized amplicons from the commercial kit. Correct genotypes were obtained at template concentrations as low as 31 pg/25 цL. Overall, our data confirm that our redesigned primers can increase the probability of obtaining a usable profile in situations where standard kits fail.

Keywords:
forensic science, DNA profiling, short tandem repeat, miniplex, degraded DNA, TH01, CSF1PO, TPOX, D5S818, D8S1179, D16S539, FGA, D21S11, D7S820, vWA, D18S51, D13S317, Penta D, Penta E, D2S1338

Paper ID: JFS2003269
DOI: 10.1520/JFS2003269
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