Recovery of genomic DNA from lingual mucosal cells in sufficient quantity and quality.
OBJECTIVE
To investigate whether genomic DNA can be purified in sufficient quantity and quality from the oral cavity.


METHODS
One milliliter of peripheral blood and saliva were collected. The buccal and lingual mucosal cells were also obtained using 10 strokes with a swab or a toothbrush, respectively. All materials were centrifuged and the cells were lysed by adding sodium dodecyl-sulfate and proteinase K. The DNAs were extracted with phenol and precipitated with ethanol followed by electrophoresing on 0.8% agarose gel. The purified DNAs were digested with restriction enzyme Dpn I and Mbo I, respectively. Amplification of the IL-1A gene by PCR was carried out using the purified DNAs and electrophoresing on polyacrylamide gel.


RESULTS
DNA was obtained from lingual mucosal cells collected with a toothbrush. Only about one-thirtieth of the recovered DNA was of non-human origin (bacterial contaminants from the oral cavity). Judging from the PCR amplifications of the IL-1A gene, the DNA extracted from lingual cells was of sufficient quality, in all respects indistinguishable from the DNAs extracted from the other specimen, such as peripheral blood, saliva and buccal mucosal cells collected with a swab, and in sufficient quantity. Our results indicate that it is possible to purify DNAs from lingual mucosal cells collected with a toothbrush in a simple and safe manner. Compared to DNA samples from patients by blood extraction, the described method also had the advantage of being painless and not inducing mental distress.
Imamura
Yasuhiro
Y
Department of Pharmacology, Matsumoto Denatal University, Nagano, Japan.
Usui
Syuhei
S
Oomori
Yuriko
Y
Fujigaki
Yoshihisa
Y
Ota
Norio
N
Wang
Pao-Li
PL
D004247:DNA / Q000097:blood / Q000235:genetics*; D016678:Genome / Q000235:genetics*; D006801:Humans; D053582:Interleukin-1alpha / Q000235:genetics; D009061:Mouth Mucosa / Q000166:cytology*; D012463:Saliva; D014059:Tongue
D053582:Interleukin-1alpha; D004247:DNA