Large-scale preparation of high-molecular weight DNA from buccal mucosa

Abstract

Four non-invasive methods of sampling DNA from buccal mucosa, simple rinses, scrubbing with cotton balls, scrubbing with toothbrushes and rinses after scrubbing with toothbrushes, were investigated. Scrubbing with toothbrushes yielded 5.79 ± 5.56 μg of DNA rich in high-molecules, while less than one eighth the amount was recovered by scrubbing with cotton balls. Rinses after scrubbing with toothbrushes gave 50.0 ± 46.0 μg of DNA and simple rinses 34.4 ± 35.7 μg, although the DNA was considerably degraded. DNA specimens obtained from buccal cells were shown to be more or less in the process of degradation including apoptosis. For minisatellite analysis, only DNA prepared by scrubbing with toothbrushes could be used, while all specimens could be applied to PCR analyses. Since scrubbing with toothbrushes is painless and harmless, we recommend this method. Subsequent rinsing will yield a large amount of DNA suitable for many PCR analyses.