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Comparison between DNA obtained from buccal cells of the upper and lower gutter area
Nedel, Fernanda; Conde, Marcus Cristian Muniz; Oliveira, Isabel Oliveira de; Tarquinio, Sandra Beatriz Chaves; Demarco, Flávio Fernando.
  • Nedel, Fernanda; Federal University of Pelotas. Dental School. Pelotas. BR
  • Conde, Marcus Cristian Muniz; Federal University of Pelotas. Dental School. Pelotas. BR
  • Oliveira, Isabel Oliveira de; Federal University of Pelotas. Institute of Biology. Department of Physiology and Pharmacology. Pelotas. BR
  • Tarquinio, Sandra Beatriz Chaves; Federal University of Pelotas. Dental School. Department of Semiology and Clinics. Pelotas. BR
  • Demarco, Flávio Fernando; Federal University of Pelotas. Dental School. Department of Restorative Dentistry. Pelotas. BR
Braz. dent. j ; 20(4): 275-278, 2009. ilus
Article in English | LILACS | ID: lil-536314
ABSTRACT
This study compared quantitatively and qualitatively the DNA extracted from buccal cells collected from the upper or lower gutter areas. Buccal cells were collected from the upper (n=15) and lower gutter (n=15) region from 15 volunteers using a special cytobrush (Gentra), totaling 2 collections from each individual. DNA was extracted from the samples according to the manufacturer's instructions. The DNA obtained was qualitatively and quantitatively evaluated by 2 calibrated blind examiners using spectrophotometry and analysis of DNA bands (0.8 percent agarose gel electrophoresis). Data was statistically analyzed by one-way ANOVA (?=0.05). Means and standard derivation (SD) for total DNA yield from the upper and lower gutter area were 12.2 ?g (12.0) and 9.4 ?g (8.5), respectively (p=0.821). There was higher (p<0.05) DNA purity for the upper gutter (1.79; 0.05) when compared to lower gutter area (1.66; 0.10). Regarding to the DNA quality, no differences were observed between the 2 location sites, but all samples showed similar degree of degradation. In conclusion, it would be recommendable that buccal cells for DNA extraction be collected from the upper gutter area in the attempt to increase DNA purity.
RESUMO
O objetivo do presente estudo foi comparar quantitativamente e qualitativamente o DNA extraído de células epiteliais bucais coletadas do fundo de sulco superior e inferior. Foram coletadas células bucais do fundo de sulco superior (n=15) e inferior (n=15) de 15 voluntários utilizando escovas citológicas especiais (Gentra), totalizando 2 coletas por voluntário. Após a coleta o DNA foi extraído conforme o protocolo indicado pelo fabricante (Puregene DNA Buccal Cell Kit; Gentra Systems, Inc.). O DNA obtido foi avaliado quantitativamente e qualitativamente por dois examinadores calibrados cegos utilizando espectrofotometria e análise das bandas de DNA (gel de agarose 0,8 por cento, por eletroforese). Os dados foram submetidos a ANOVA a um critério, com p<0,05. As médias e desvio padrão (DP) para o rendimento total de DNA do fundo de sulco superior e inferior foram respectivamente 12,2 ?g (12,0) e 9,4 ?g (8,5) (p=0,821). Houve maior (p<0,05) pureza de DNA no fundo de sulco superior (1,79; 0,05) quando comparado com o fundo de sulco inferior (1,66; 0,10). Quanto à qualidade do DNA, não foi observado diferenças entre os dois locais testados, no entanto todas as amostras mostraram níveis de degradação semelhantes. Em conclusão seria recomendável coletar células bucais, para extração de DNA, do fundo de sulco superior na tentativa de aumentar a pureza do DNA.
Subject(s)

Full text: Available Index: LILACS (Americas) Main subject: Specimen Handling / In Vitro Techniques / DNA / Mouth Mucosa Type of study: Controlled clinical trial / Evaluation studies Limits: Adolescent / Adult / Humans Language: English Journal: Braz. dent. j Journal subject: Dentistry Year: 2009 Type: Article Affiliation country: Brazil Institution/Affiliation country: Federal University of Pelotas/BR

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Full text: Available Index: LILACS (Americas) Main subject: Specimen Handling / In Vitro Techniques / DNA / Mouth Mucosa Type of study: Controlled clinical trial / Evaluation studies Limits: Adolescent / Adult / Humans Language: English Journal: Braz. dent. j Journal subject: Dentistry Year: 2009 Type: Article Affiliation country: Brazil Institution/Affiliation country: Federal University of Pelotas/BR