title: Rapid extraction and preservation of genomic DNA from human samples.
abstract: Simple and rapid extraction of human genomic DNA remains a bottleneck 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 onto 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 four 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 1 month was demonstrated for captured DNA, facilitating straightforward collection, delivery, and handling of genomic DNA in an environment-friendly protocol.
authors.affiliation: Department of Mechanical Engineering, University of Washington, Seattle, WA 98195, USA.
authors.forename: D
authors.initials: D
authors.lastname: Kalyanasundaram
authors.forename: J-H
authors.initials: JH
authors.lastname: Kim
authors.forename: W-H
authors.initials: WH
authors.lastname: Yeo
authors.forename: K
authors.initials: K
authors.lastname: Oh
authors.forename: K-H
authors.initials: KH
authors.lastname: Lee
authors.forename: M-H
authors.initials: MH
authors.lastname: Kim
authors.forename: S-M
authors.initials: SM
authors.lastname: Ryew
authors.forename: S-G
authors.initials: SG
authors.lastname: Ahn
authors.forename: D
authors.initials: D
authors.lastname: Gao
authors.forename: G A
authors.initials: GA
authors.lastname: Cangelosi
authors.forename: J-H
authors.initials: JH
authors.lastname: Chung
mesh_terms: D002467:Cell Nucleus / Q000737:chemistry; D004247:DNA / Q000302:isolation & purification*; D004272:DNA, Mitochondrial / Q000302:isolation & purification*; D055664:Electrochemical Techniques; D006046:Gold / Q000737:chemistry; D006801:Humans; D046208:Microchip Analytical Procedures / Q000592:standards; D009061:Mouth Mucosa / Q000737:chemistry*; D009691:Nucleic Acid Denaturation; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation* / Q000379:methods; D016133:Polymerase Chain Reaction; D011309:Preservation, Biological / Q000379:methods*; D012463:Saliva / Q000737:chemistry*; D013048:Specimen Handling / Q000295:instrumentation / Q000379:methods
chemical_list: D004272:DNA, Mitochondrial; D006046:Gold; D004247:DNA