Evaluation and Quantification of Nuclear DNA from Human Telogen Hairs

Department of Chemistry and Biochemistry, Florida International University, University Park, 11200 SW 8th Street, Miami, FL 33199, USA.
Journal of Forensic Sciences (Impact Factor: 1.16). 07/2008; 53(4):853-7. DOI: 10.1111/j.1556-4029.2008.00777.x
Source: PubMed


Nuclear DNA was extracted from human telogen hairs from 60 individuals. Six to nine hairs from each individual were individually extracted. The amount of DNA recovered from each individual varied greatly, and most samples yielded a quantity of 550 pg or less per hair. A selective extraction buffer was used to remove epithelial cell DNA and the amount of exogenous DNA was determined. DNA was also quantified by real time PCR using three different sized amplicons targeting an Alu sequence. The results were used to determine the state of degradation of the extracted DNA. Different quantities of sample (<100 pg, 100-500 pg, >500 pg) were amplified with the Miniplex kits to determine the minimum DNA template required for successful amplification. DNA recovered from hair showed degradation; however, partial profiles were obtained for those samples containing at least 60 pg using MiniSTRs.

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Available from: Bruce Mccord, Mar 31, 2014
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    • "Quantitative real-time PCR (qPCR) was performed using the LightCycler® Carousel-Based System 1.2 and the Light Cycler® Fast Start DNA Master SYBR Green I according to the manufacturer’s protocol (Roche Applied Science). Nuclear DNA was amplified with the primers ALU-forward 5′-GTCAGGAGATCGAGACCATCCC-3′ and 124-bp-reverse 5′-TCCTGCCTCAGCCTCCCAAG-3′ [8]. Mitochondrial DNA was amplified with the primers mito-for 5′-CTCAGATAGGGGTCCCTTGA-3′ and mito-rev 5′-GCACTCTTGTGCGGGATATT-3′ which align to nucleotides 16380–16399 and 16420–16439, respectively of the human mitochondrial genome (GenBank accession number NC_012920.1). "
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