Production and certification of NIST Standard Reference Material 2372 Human DNA Quantitation Standard |
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Authors: | Margaret C Kline David L Duewer John C Travis Melody V Smith Janette W Redman Peter M Vallone Amy E Decker and John M Butler |
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Institution: | (1) Biochemical Science Division, National Institute of Standards and Technology, 100 Bureau Drive, MS 8311, Gaithersburg, MD 20899-8311, USA;(2) Analytical Chemistry Division, National Institute of Standards and Technology, 100 Bureau Drive, MS 8390, Gaithersburg, MD 20899-8390, USA |
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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.
Figure NIST Standard Reference Material (SRM) 2372 Human Quantitation Standard |
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Keywords: | Certified reference material (CRM) Decadic attenuance Forensic Human identity Interlaboratory comparison Short tandem repeat (STR) multiplex assay Standard Reference Material (SRM) UV/visible absorbance spectrophotometry |
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