Abstract

Anonymous liquid blood samples with self-identified ethnicities were purchased from Interstate Blood Bank (Memphis, TN) and Millennium Biotech, Inc. (Ft. Lauderdale, FL) and extracted using a modified salt out procedure (1). The extracted DNA was then quantified using UV spectrophotometry at 260 nm and a PicoGreen assay (2). A 150-µL aliquot of the extracted DNA solution was directly quantified in a Cary 100 double-beam spectrophotometer (Varian Analytical Instruments, Walnut Creek, CA). Low volume micro-cuvettes allowed for accurate absorbance measurements (A = 0.2 to 0.6) without prior dilution of the stock extracted DNA. Sample concentrations were adjusted to 1 ng/µL for typing purposes using the PicoGreen assay values. Fifteen autosomal STR markers (the 13 CODIS core loci and D19S433 and D2S1338) were typed along with amelogenin using the Applied Biosystems AmpF1STR® Identifiler™ kit (3). PCR amplification was carried out on a GeneAmp® 9700 (Applied Biosystems) using 1 ng of DNA according to kit protocols (3) with the exception of reduced volume reactions (5 µL instead of 25 µL) and reduced cycles (26 instead of 28). Amplification products were diluted 1:15 in HiDi™ formamide and GS500-LIZ internal size standard (Applied Biosystems) and analyzed on the 16-capillary ABI Prism® 3100 Genetic Analyzer without prior denaturation of samples. POP™-6 (Applied Biosystems) rather than POP™-4 was utilized for higher resolution separations on a 36 cm array. Samples were injected

Author Information

Butler, JM
Schoske, R
National Institute of Standards and Technology, Gaithersburg, MD
Vallone, PM
National Institute of Standards and Technology, Gaithersburg, MD
Redman, JW
National Institute of Standards and Technology, Gaithersburg, MD
Kline, MC
Pages: 4
Price: $25.00
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Stock #: JFS2003045
ISSN: 0022-1198
DOI: 10.1520/JFS2003045