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Proceedings Paper

Small sample whole-genome amplification
Author(s): Christine Hara; Christine Nguyen; Elizabeth Wheeler; Karen Sorensen; Erin Arroyo; Greg Vrankovich; Allen Christian
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Paper Abstract

Many challenges arise when trying to amplify and analyze human samples collected in the field due to limitations in sample quantity, and contamination of the starting material. Tests such as DNA fingerprinting and mitochondrial typing require a certain sample size and are carried out in large volume reactions; in cases where insufficient sample is present whole genome amplification (WGA) can be used. WGA allows very small quantities of DNA to be amplified in a way that enables subsequent DNA-based tests to be performed. A limiting step to WGA is sample preparation. To minimize the necessary sample size, we have developed two modifications of WGA: the first allows for an increase in amplified product from small, nanoscale, purified samples with the use of carrier DNA while the second is a single-step method for cleaning and amplifying samples all in one column. Conventional DNA cleanup involves binding the DNA to silica, washing away impurities, and then releasing the DNA for subsequent testing. We have eliminated losses associated with incomplete sample release, thereby decreasing the required amount of starting template for DNA testing. Both techniques address the limitations of sample size by providing ample copies of genomic samples. Carrier DNA, included in our WGA reactions, can be used when amplifying samples with the standard purification method, or can be used in conjunction with our single-step DNA purification technique to potentially further decrease the amount of starting sample necessary for future forensic DNA-based assays.

Paper Details

Date Published: 11 November 2005
PDF: 5 pages
Proc. SPIE 6007, Smart Medical and Biomedical Sensor Technology III, 600717 (11 November 2005); doi: 10.1117/12.630925
Show Author Affiliations
Christine Hara, Lawrence Livermore National Lab. (United States)
Christine Nguyen, Lawrence Livermore National Lab. (United States)
Elizabeth Wheeler, Lawrence Livermore National Lab. (United States)
Karen Sorensen, Lawrence Livermore National Lab. (United States)
Erin Arroyo, Lawrence Livermore National Lab. (United States)
Greg Vrankovich, Lawrence Livermore National Lab. (United States)
Allen Christian, Lawrence Livermore National Lab. (United States)


Published in SPIE Proceedings Vol. 6007:
Smart Medical and Biomedical Sensor Technology III
Brian M. Cullum; J. Chance Carter, Editor(s)

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