Method to Determine the 5' End of the Binding Site of Primers Included in a Commercially Available Forensic Human Identification Kit

• NCJ Number: 245537
• Journal: Forensic Science International: Genetics Volume: 9 Dated: March 2014 Pages: 76-80
• Author(s): Natsuko Mizuno; Shota Inokuchi; Tetsushi Kitayama; Koji Fujii; Kentaro Kasai; Kazumasa Sekiguchi
• Date Published: March 2014
• Length: 5 pages
• Publication Format: Article
• Publication Type: Research (Applied/Empirical); Report (Study/Research)
• Language: English
• Country: Ireland

Annotation

This study developed a simple method for determining the 5' end of the binding site of the primers included in commercial forensic human identification kits, since the manufacturers of the majority of such kits do not disclose the sequences of the primers included in the kit.

Abstract

The proposed method requires custom primers and human genome sequence data, as well as equipment and consumables routinely used in laboratories. One or two custom primers are added to the PCR reaction mixture that contains kit primers and input human DNA prior to amplification. PCR products are separated by capillary electrophoresis after amplification. This method can determine which primer of the pair is fluorescently labeled and the 5' end of the binding site of primers based on the changes in an electropherogram that are caused by the addition of the custom primer(s), as well as the human genome sequence data. This method can also be used to determine the shortest possible lengths of labeled kit primers. The detailed description of materials and methods addresses the features of the primers and PCR amplification. 5 figures, 2 tables, and 14 references