DNA Quantitation Efforts by the NIST Forensics/Human Identity Project Team

Team Members: John M. Butler (Project Leader), Margaret C. Kline, Peter M. Vallone, Janette W. Redman, Amy E. Decker,  Carolyn R. "Becky" Hill, David L. Duewer (Analytical Chemistry Division)

$Funding from the National Institute of Justice through the NIST Office of Law Enforcement Standards

Real-time quantitative PCR (qPCR) has generated a great deal of interest in the forensic DNA typing community in the past several years as this technique can rapidly detect low levels of DNA with minimum hands-on time and minor sample consumption. The ability to utilize human specific assays in U.S. crime laboratories is important in order to meet federally mandated requirements (DAB/FBI Standard 9.3) to assess the quantity of human DNA in casework samples, particularly where bacterial contamination may exist.

Poster describing some NIST work regarding issues behind real-time qPCR for human DNA quantitation

Kline, M.C., Vallone, P.M., Decker, A.E., Redman, J.W., Duewer, D.L., Butler, J.M. (2005) Testing candidate DNA quantitation standards with several real-time quantitative PCR methods. Proceedings of the 16^th International Symposium on Human Identification. http://www.promega.com/geneticidproc/ussymp16proc/abstracts/kline.pdf.

Margaret Kline talk at the 16th International Symposium on Human Identification (Grapevine, TX), September 29, 2005, "Testing Candidate DNA Quantitation Standards with Several Real-Time Quantitative PCR Methods" [.pdf]

Peter Vallone talk at 58th Annual Meeting of the American Academy of Forensic Sciences (Seattle, WA), February 23, 2006, "Examining Candidate DNA Quantitation Standards with Real-Time Quantitative PCR Assays" [.pdf]

Margaret Kline presentation at the NIJ DNA Grantees meeting (Crystal City, VA), June 26, 2006, "Progress Toward SRM 2372: Human DNA Quantitation" [.pdf]

Peter Vallone qPCR workshop with Cristian Orrego held at the National Forensic Science Technology Center (Largo, FL), July 26-27, 2006 [Introduction] [Quantitation Using PCR] [Instrumentation] [Signals and Probes] Selected Forensic qPCR Assays: [VT Alu SYBR Green and Quantifiler] [CA DOJ nuclear/mtDNA duplex and CA DOJ degradation triplex] [Maintenance, Validation, etc.] [Data Analysis and Troubleshooting] [qPCR Analysis Software for ABI 7000 & 7500]

Margaret Kline presentation at the Mid-Atlantic Association of Forensic Scientists annual meeting (Washington, DC), May 25, 2007, "SRM 2372: How the Human DNA Quantitation Standard was Characterized at NIST and How It Can Be Used in Your Laboratory" [.pdf]

More information coming soon...