DNA from buccal swabs suitable for high-throughput SNP multiplex analysis.
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 microg +/- 0.54 with a 94% genotyping pass rate. For the high-throughput application (n=1140), the average DNA yield was 2.44 microg +/- 1.74 with a >or=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.
McMichael
Gai L
GL
The University of Adelaide, Disciplines of Obstetrics and Gynaecology, Women's and Children's Hospital, Adelaide, Australia. gai.mcmichael@adelaide.edu.au
Gibson
Catherine S
CS
O'Callaghan
Michael E
ME
Goldwater
Paul N
PN
Dekker
Gustaaf A
GA
Haan
Eric A
EA
MacLennan
Alastair H
AH
D000293:Adolescent; D000328:Adult; D000368:Aged; D001331:Automation; D020811:Biomedical Technology / Q000379:methods*; D002610:Cheek; D002648:Child; D015331:Cohort Studies; D004247:DNA / Q000378:metabolism*; D005838:Genotype; D006801:Humans; D008875:Middle Aged; D008957:Models, Genetic; D010865:Pilot Projects; D020641:Polymorphism, Single Nucleotide
D004247:DNA