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Anal Biochem. 2016 Feb 1;494:40-5. doi: 10.1016/j.ab.2015.10.002. Epub 2015 Oct 22.

High-resolution melt analysis of DNA methylation to discriminate semen in biological stains.

Author information

1
Department of Chemistry and Biochemistry, Florida International University, Miami, FL 33199, USA.
2
Department of Chemistry and Biochemistry, Florida International University, Miami, FL 33199, USA; Faculty of Biosciences, Laboratory of Human and Molecular Genetics, PUCRS, 90619-900 Porto Alegre, Brazil.
3
School of Criminal Justice, University of Southern Mississippi, Hattiesburg, MS 39406, USA.
4
DNA Laboratory, Broward County Sheriff's Office, Fort Lauderdale, FL 33301, USA.
5
Faculty of Biosciences, Laboratory of Human and Molecular Genetics, PUCRS, 90619-900 Porto Alegre, Brazil.
6
Department of Chemistry and Biochemistry, Florida International University, Miami, FL 33199, USA. Electronic address: mccordb@fiu.edu.

Abstract

The goal of this study was to develop a method for the detection of semen in biological stains using high-resolution melt (HRM) analysis and DNA methylation. To perform this task, we used an epigenetic locus that targets a tissue-specific differentially methylated region for semen. This specific locus, ZC3H12D, contains methylated CpG sites that are hypomethylated in semen and hypermethylated in blood and saliva. Using this procedure, DNA from forensic stains can be isolated, processed using bisulfite-modified polymerase chain reaction (PCR), and detected by real-time PCR with HRM capability. The method described in this article is robust; we were able to obtain results from samples with as little as 1 ng of genomic DNA. Samples inhibited by humic acid still produced reliable results. Furthermore, the procedure is specific and will not amplify non-bisulfite-modified DNA. Because this process can be performed using real-time PCR and is quantitative, it fits nicely within the workflow of current forensic DNA laboratories. As a result, it should prove to be a useful technique for processing trace evidence samples for serological analysis.

KEYWORDS:

Body fluid identification; DNA methylation; Epigenetic; Forensic; High-resolution melt; Real-time PCR

PMID:
26470939
DOI:
10.1016/j.ab.2015.10.002
[Indexed for MEDLINE]

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