Forensic DNA: Human DNA Quantitation

Description

When biological evidence from a crime scene is processed to isolate the DNA present, all sources of DNA are extracted. Thus, non-human DNA such as bacterial, fungal, plant, or animal material may also be present in the total DNA recovered from the sample along with the relevant human DNA of interest. For this reason, the DNA Advisory Board (DAB) Standards that govern forensic DNA testing of forensic casework require human-specific DNA quantitation (standard 9.3). This requirement ensures that appropriate levels of human DNA can be included in the subsequent polymerase chain reaction (PCR) amplification of short tandem repeats (STRs) evaluated in a DNA profile.

Equally important is the fact that multiplex STR typing works best with a fairly narrow range of human DNA. Typically 0.5 to 2.0 ng of input DNA works best with commercial STR kits. Too much DNA results in overblown electropherograms that make interpretation of results more challenging. Too little DNA can result in loss of alleles due to stochastic amplification in a low copy number regime.

In recent years, research in human DNA quantitation has focused on new "real-time" quantitative PCR (qPCR) techniques. Quantitative PCR methods enable automated, precise, and high-throughput measurements. Interlaboratory studies have demonstrated the importance of human DNA quantitation on achieving reliable interpretation of STR typing and obtaining consistent results across laboratories.

Research Projects

This table below present awards funded by the National Institute of Justice both prior to and under the DNA Initiative, which began in 2004.

TitleAwardeeAmountAward Number
Population Genetic Issues for Forensic DNA ProfilesUniversity of Washington2011-DN-BX-K541$1,269,456
Quantitation of DNA for Forensic DNA Typing by qPCRCalifornia Department of Justice2002-IJ-CX-K008$126,821
Simple, Rapid, and Accurate Quantitation of Human DNAVermont Department of Public Safety2000-IJ-CX-K012$67,129
Simple, Rapid, and Accurate Quantitation of Human DNAVermont Department of Public Safety2000-IJ-CX-K012$124,970
Simple, Rapid, and Accurate Quantitation of Human DNAVermont Department of Public Safety2000-IJ-CX-K012$192,033

Research Articles and Reports from Funded Projects

Select a title to view an abstract from PubMed database (National Library of Medicine's) unless a different file type is indicated.

TitleAuthorsPublicationYearAwardeeAward Number
Production and certification of NIST Standard Reference Material 2372 Human DNA Quantitation Standard. Kline MC, Duewer DL, Travis JC, Smith MV, Redman JW, Vallone PM, Decker AE, Butler JM.Anal Bioanal Chem. 2009 Apr 18. [Epub ahead of print]2009NIST2003-IJ-R-029
A quadruplex real-time qPCR assay for the simultaneous assessment of total human DNA, human male DNA, DNA degradation and the presence of PCR inhibitors in forensic samples: A diagnostic tool for STR typingWilliam R. Hudlow, Mavis Date Chong, Katie L. Swango,
Mark D. Timken, Martin R. Buoncristiani
Forensic Science International: Genetics 2 (2008) 108–1252008California Department of Justice2002-IJ-CX-K008
Developmental validation of a multiplex qPCR assay for assessing the quantity and quality of nuclear DNA in forensic samples.Swango KL, Hudlow WR, Timken MD, Buoncristiani MR.Forensic Sci Int. 2006 Oct 27; [Epub ahead of print]2006California Department of Justice2002-IJ-CX-K008
Simultaneous determination of total human and male DNA using a duplex real-time PCR assay.Nicklas JA and Buel E.Journal of Forensic Sciences 2006 September; 51(5): 1005-1015.2006Vermont Department of Public Safety2000-IJ-CX-K012
Simple, Rapid and Accurate Quantitation of Human DNA (pdf, 101 pages)Eric Buel, Ph.D.Final Grant Report2006Vermont Department of Public Safety2000-IJ-CX-K012
A duplex real-time qPCR assay for the quantification of human nuclear and mitochondrial DNA in forensic samples: Implications for quantifying DNA in degraded samples. Timken MD, Swango KL, Orrego C, Buoncristiani MRJ Forensic Sci. 2005 Sep; 50(5):1044-1060.2005California Department of Justice2002-IJ-CX-K008
Quantitation of DNA for Forensic DNA Typing by qPCR (pdf, 90 pages)Mark D. Timken, Katie L. Swango, Cristián Orrego, Mavis Date Chong, Martin R. BuoncristianiFinal Grant Report2005California Department of Justice2002-IJ-CX-K008
An Alu-based, MCG Eclipsetm Real-time PCR method for quantitation of human DNA in forensic samplesNicklas JA, Buel EJournal of Forensic Science. 2005 Sep; 50(5):1081-10902005Vermont Department of Public Safety2000-IJ-CX-K012
Results from the NIST 2004 DNA Quantitation Study.Kline MC, Duewer DL, Redman JW, Butler JM.J Forensic Sci. 2005 May; 50(3):570-8.2005National Institute of Standards and Technology2003-IJ-R-029
Development of an Alu-based, QSY 7-labeled primer PCR method for quantitation of human DNA in forensic samples.Nicklas JA, Buel E.Journal of Forensic Science 2003 Mar; 48(2):282-91.2003Vermont Department of Public Safety2000-IJ-CX-K012
Development of an Alu-based, real-time PCR method for quantitation of human DNA in forensic samples.Nicklas JA, Buel E.Journal of Forensic Science. 2003 Sep; 48(5):936-44.2003Vermont Department of Public Safety2000-IJ-CX-K012
Quantification of DNA in forensic samples.Nicklas JA, Buel E.Anal Bioanal Chem. 2003 Aug; 376(8):1160-7. Epub 2003 May 09.2003Vermont Department of Public Safety2000-IJ-CX-K012
Date Modified: October 11, 2012