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                            Library & Multimedia
 Production and certification of NIST Standard Reference Material 2372 Human DNA Quantitation Standard
NCJ Number
              308352
Journal

*Analytical and Bioanalytical Chemistry* Volume: 394 Dated: 2009 Pages: 1183–1192
Author(s)
          Margaret C. Kline;
                        David L. Duewer;
                        John C. Travis;
                        Melody V. Smith;
                        Janette W. Redman;
                        Peter M. Vallone;
                        Amy E. Decker;
                        John M. Butler
Date Published

2009
Length
              10 pages
Annotation
              This paper discusses the practicality of common methods used to quantitate stable human DNA, in order to provide a certified reference material that will help the forensic community reduce within- and among-laboratory quantitative variability.
Abstract
              Modern highly multiplexed short tandem repeat (STR) assays used by the forensic human-identity community require tight control of the initial amount of sample DNA amplified in the polymerase chain reaction (PCR) process. This, in turn, requires the ability to reproducibly measure the concentration of human DNA, [DNA] in a sample extract. Quantitative PCR (qPCR) techniques can determine the number of intact stretches of DNA of specified nucleotide sequence in an extremely small sample; however, these assays must be calibrated with DNA extracts of well-characterized and stable composition. By 2004, studies coordinated by or reported to the National Institute of Standards and Technology (NIST) indicated that a well-characterized, stable human DNA quantitation certified reference material (CRM) could help the forensic community reduce within- and among-laboratory quantitation variability. To ensure that the stability of such a quantitation standard can be monitored and that, if and when required, equivalent replacement materials can be prepared, a measurement of some stable quantity directly related to [DNA] is required. Using a long-established conventional relationship linking optical density (properly designated as decadic attenuance) at 260 nm with [DNA] in aqueous solution, NIST Standard Reference Material (SRM) 2372 Human DNA Quantitation Standard was issued in October 2007. This SRM consists of three quite different DNA extracts: a single-source male, a multiple-source female, and a mixture of male and female sources. All three SRM components have very similar optical densities, and thus very similar conventional [DNA]. The materials perform very similarly in several widely used gender-neutral assays, demonstrating that the combination of appropriate preparation methods and metrologically sound spectrophotometric measurements enables the preparation and certification of quantitation [DNA] standards that are both maintainable and of practical utility. (Published Abstract Provided)
      
  
          Additional Details
Grant Number(s)
        2008-DN-R-121
                                2003-IJ-R-029


  
  
        
  Agencies
        NIJ-Sponsored


  
  
        
  Publication Type
        Research (Applied/Empirical)
    
  
  
Date Published: January 1, 2009
    
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