DNA from Buccal Swabs Suitable for High-Throughput SNP Multiplex Analysis
Date
2009
Authors
McMichael, G.
Gibson, C.
O'Callaghan, M.
Goldwater, P.
Dekker, G.
Haan, E.
MacLennan, A.
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Advisors
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Journal article
Citation
Journal of Biomolecular Techniques, 2009; 20(5):232-235
Statement of Responsibility
Gai L. McMichael, Catherine S. Gibson, Michael E. O’Callaghan, Paul N. Goldwater, Gustaaf A. Dekker, Eric A. Haan, Alastair H. MacLennan, and for the South Australian Cerebral Palsy Research Group
Conference Name
Abstract
We sought a convenient and reliable method for collection of genetic material that is inexpensive and noninvasive and suitable for self-collection and mailing and a compatible, commercial DNA extraction protocol to meet quantitative and qualitative requirements for high-throughput single nucleotide polymorphism (SNP) multiplex analysis on an automated platform. Buccal swabs were collected from 34 individuals as part of a pilot study to test commercially available buccal swabs and DNA extraction kits. DNA was quantified on a spectrofluorometer with Picogreen dsDNA prior to testing the DNA integrity with predesigned SNP multiplex assays. Based on the pilot study results, the Catch-All swabs and Isohelix buccal DNA isolation kit were selected for our high-throughput application and extended to a further 1140 samples as part of a large cohort study. The average DNA yield in the pilot study (n=34) was 1.94 μg ± 0.54 with a 94% genotyping pass rate. For the high-throughput application (n=1140), the average DNA yield was 2.44 μg ± 1.74 with a ≥93% genotyping pass rate. The Catch-All buccal swabs are a convenient and cost-effective alternative to blood sampling. Combined with the Isohelix buccal DNA isolation kit, they provided DNA of sufficient quantity and quality for high-throughput SNP multiplex analysis.
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Description
Copyright © 2009 Association of Biomolecular Resource Facilities