Rapid extraction and preservation of genomic DNA from human samples

• Published in: Analytical and bioanalytical chemistry Vol. 405; no. 6; pp. 1977 - 1983
• Main Authors: Kalyanasundaram, D., Kim, J.-H., Yeo, W.-H., Oh, K., Lee, K.-H., Kim, M.-H., Ryew, S.-M., Ahn, S.-G., Gao, D., Cangelosi, G. A., Chung, J.-H.
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Springer-Verlag 01.02.2013; Springer; Springer Nature B.V
• Subjects: Analytical Chemistry; Biochemistry; Cell Nucleus - chemistry; Characterization and Evaluation of Materials; Charged particles; Chemistry; Chemistry and Materials Science; Deoxyribonucleic acid; DNA; DNA - isolation & purification; DNA, Mitochondrial - isolation & purification; Electric fields; Electrochemical Techniques; Extraction; Extraction (Chemistry); Food Science; Genetic testing; Genomes; Genomics; Gold - chemistry; Handling; Human; Human genetics; Humans; Laboratory Medicine; Materials handling; Methods; Microchip Analytical Procedures - standards; Monitoring/Environmental Analysis; Mouth Mucosa - chemistry; Nucleic Acid Denaturation; Oligonucleotide Array Sequence Analysis - instrumentation; Oligonucleotide Array Sequence Analysis - methods; Original Paper; Physiological aspects; Polymerase Chain Reaction; Preservation; Preservation, Biological - methods; Protocol; Saliva - chemistry; Semiconductors; Silicon nitride; Specimen Handling - instrumentation; Specimen Handling - methods; United States; United States > US; DNA extraction; Human genomic DNA; Electric field; Microtip; Human samples
• ISSN: 1618-2642; 1618-2650
• DOI: 10.1007/s00216-012-6637-8

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. Figure Portable microtip device for human genomic DNA extraction