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.
From research sponsored by the National Institute of Justice
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Simple, Rapid, and Accurate Quantitation of Human DNA
Vermont Department of Public Safety
Award Number: 2000-IJ-CX-K012
Final Report (pdf)Quantitation of DNA for Forensic DNA Typing by qPCR: Singleplex and Multiplex Modes for Nuclear and Mitochondrial Genomes, and the Y Chromosome
California Department of Justice
Award Number: 2002-IJ-CX-K008
Final Report (pdf)
From research sponsored by the National Institute of Justice
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.
View Pub Med EntryGrant number: 2000-IJ-CX-K012
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]
View Pub Med EntryGrant number: 2002-IJ-CX-K008
A quantitative PCR assay for the assessment of DNA degradation in forensic samples.
Swango KL, Timken MD, Chong MD, Buoncristiani MR.
Forensic Sci Int. 2006 Apr 20;158(1):14-26. Epub 2005 Jun 3
View Pub Med Entry
Grant number: 2002-IJ-CX-K008
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.
Grant number: 2003-IJ-R-029
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 MR
J Forensic Sci. 2005 Sep;50(5):1044-1060.
View Pub Med Entry Grant number: 2002-IJ-CX-K008 Swango KL, Timken MD, Chong MD, Buoncristiani MR.
An Alu-based, MCG Eclipsetm Real-time PCR method for quantitation of human DNA in forensic samples Nicklas JA, Buel E
Journal of Forensic Science, 2005 Sep;50(5):1081-1090.
View Pub Med Entry Grant number: 2000-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.
View Pub Med Entry Grant number: 2000-IJ-CX-K012
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.
View Pub Med Entry Grant number: 2000-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.
View Pub Med Entry
Grant number: 2000-IJ-CX-K012