Biology Faculty Articles
Title
High-resolution melt analysis of DNA methylation to discriminate semen in biological stains
Document Type
Article
Publication Date
2-1-2016
Publication Title
Analytical Biochemistry
Keywords
Body fluid identification, Forensic, DNA methylation, High-resolution melt, Real-time PCR, Epigenetic
ISSN
0003-2697
Volume
494
First Page
40
Last Page
45
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.
NSUWorks Citation
Antunes, Joana; Deborah S. B. S. Silva; Kuppareddi Balamurugan; George Duncan; Clarice S. Alho; and Bruce McCord. 2016. "High-resolution melt analysis of DNA methylation to discriminate semen in biological stains." Analytical Biochemistry 494, (): 40-45. doi:10.1016/j.ab.2015.10.002.
DOI
10.1016/j.ab.2015.10.002
