Accepted manuscript
Rapid extraction and preservation of genomic DNA from human samples
Analytical and bioanalytical chemistry, Vol.405(6), pp.1977-1983
02/2013
Handle:
https://hdl.handle.net/2376/112838
PMCID: PMC3566295
PMID: 23307121
Abstract
Simple and rapid extraction of human genomic DNA remains a bottle neck for genome analysis and disease diagnosis. Current methods using microfilters require cumbersome, multiple handling steps in part because salt conditions must be controlled for attraction and elution of DNA in porous silica. We report a novel extraction method of human genomic DNA from buccal swab- and saliva samples. DNA is attracted on to a gold-coated microchip by an electric field and capillary action while the captured DNA is eluted by thermal heating at 70 °C. A prototype device was designed to handle 4 microchips, and a compatible protocol was developed. The extracted DNA using microchips was characterized by qPCR for different sample volumes, using different lengths of PCR amplicon, and nuclear and mitochondrial genes. In comparison with a commercial kit, an equivalent yield of DNA extraction was achieved with fewer steps. Room-temperature preservation for one month was demonstrated for captured DNA, facilitating straightforward collection, delivery and handling of genomic DNA in an environment-friendly protocol.
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Details
- Title
- Rapid extraction and preservation of genomic DNA from human samples
- Creators
- D Kalyanasundaram - Department of Mechanical Engineering, University of Washington, Seattle, WA 98195, USAJ.-H Kim - Department of Mechanical Engineering, University of Washington, Seattle, WA 98195, USAW.-H Yeo - Department of Mechanical Engineering, University of Washington, Seattle, WA 98195, USAK Oh - NanoFacture, Inc., P.O. Box 52651, Bellevue, WA 98015, USAK.-H. LeeS.-M Ryew - KNR systems, Inc., Yongin-si, Gyeonggi-do, 449-881, Republic of KoreaS.-G Ahn - Department of Industrial Design, University of Washington, Seattle, WA 98195, USAD Gao - Department of Mechanical Engineering, University of Washington, Seattle, WA 98195, USAG. A Cangelosi - Washington Department of Environmental and Occupational Health Sciences, University of Washington, Seattle, WA 98195, USAJ.-H Chung - Department of Mechanical Engineering, University of Washington, Seattle, WA 98195, USA
- Publication Details
- Analytical and bioanalytical chemistry, Vol.405(6), pp.1977-1983
- Academic Unit
- School of Engineering and Computer Science (VANC)
- Publisher
- Springer Nature
- Number of pages
- 7
- Grant note
- 0956876 / NSF STTR II award; National Science Foundation (NSF); NSF - Directorate for Engineering (ENG) R43GM099347 / NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA; NIH National Institute of General Medical Sciences (NIGMS) NIH/NIGMS 1R43GM099347 / NIH SBIR; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA ECCS-0846454 / NSF; National Science Foundation (NSF)
- Identifiers
- 99900548243701842
- Language
- English
- Resource Type
- Accepted manuscript