Small sample whole-genome amplification

Christine Hara; Christine Nguyen; Elizabeth Wheeler; Karen Sorensen; Erin Arroyo; Greg Vrankovich; Allen Christian

doi:10.1117/12.630925

11 November 2005 Small sample whole-genome amplification

Christine Hara, Christine Nguyen, Elizabeth Wheeler, Karen Sorensen, Erin Arroyo, Greg Vrankovich, Allen Christian

Proceedings Volume 6007, Smart Medical and Biomedical Sensor Technology III; 600717 (2005) https://doi.org/10.1117/12.630925
Event: Optics East 2005, 2005, Boston, MA, United States

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.

Citation Download Citation

Christine Hara, Christine Nguyen, Elizabeth Wheeler, Karen Sorensen, Erin Arroyo, Greg Vrankovich, and Allen Christian "Small sample whole-genome amplification", Proc. SPIE 6007, Smart Medical and Biomedical Sensor Technology III, 600717 (11 November 2005); https://doi.org/10.1117/12.630925

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