Polymerase Chain Reaction Amplification of DNA from Aged Blood Stains:  Quantitative Evaluation of the “Suitability for Purpose” of Four Filter Papers as Archival Media

Abstract

In collaboration with the Armed Forces Institute of Pathology's Department of Defense DNA Registry, the National Institute of Standards and Technology recently evaluated the performance of a short tandem repeat multiplex with dried whole blood stains on four different commercially available identification card matrixes. DNA from 70 stains that had been stored for 19 months at ambient temperature was extracted or directly amplified and then processed using routine methods. All four storage media provided fully typeable (qualitatively identical) samples. After standardization, the average among-locus fluorescence intensity (electropherographic peak height or area) provided a suitable metric for quantitative analysis of the relative amounts of amplifiable DNA in an archived sample. The amounts of DNA in Chelex extracts from stains on two untreated high-purity cotton linter pulp papers and a paper treated with a DNA-binding coating were essentially identical. Average intensities for the aqueous extracts from a paper treated with a DNA-releasing coating were somewhat lower but also somewhat less variable than for the Chelex extracts. Average intensities of directly amplified punches of the DNA-binding paper were much larger but somewhat more variable than the Chelex extracts. Approximately 25% of the observed variation among the intensity measurements is shared among the four media and thus can be attributed to intrinsic variation in white blood count among the donors. All of the evaluated media adequately "bank" forensically useful DNA in well-dried whole blood stains for at least 19 months at ambient temperature.

Zinc supplementation of young men alters metallothionein, zinc transporter, and cytokine gene expression in leukocyte populations

“…Furthermore, dried blood spots (DBS) extracted from collection cards have routinely been used for DNA amplification for population screening, e.g., genotyping and for DNA archiving (5,6), but have not been widely used for RT-PCR (7)(8)(9). Such an approach of sample acquisition could similarly be used to measure transcript abundance for nutritionally regulated genes, and, therefore, be of value for nutrient assessment purposes.…”

Forensic DNA Analysis

“…For this sample type, method comparisons have not found any significant differences in performance (Viltrop et al 2010). Liquid blood and saliva are often transferred to filter paper for storage, and some brands will trap the DNA in the paper, which means any inhibitors can be removed in a few wash steps and the cutting can be added to the PCR reaction (Kline et al 2002). New formulations for the PCR buffer have made it possible to omit these initial purification steps and place any cutting or robotic punch of any storage card in a direct PCR (Park et al 2008;Wang et al 2011).…”

Setting standards and developing technology to aid the human identity testing community

“…Over the past decade, a number of studies have been performed to examine DNA stability on various solid matrices including untreated S&S 903 paper, Whatman, IsoCode, and FTA treated papers [37]. We have been able to successfully recover full STR profiles from bloodstains stored on untreated paper for 15 years at room temperature.…”