title: Design of a protocol for obtaining genomic DNA from saliva using mouthwash: Samples taken from patients with periodontal disease.
abstract: BACKGROUND
  Obtaining high quality genomic DNA safely and economically is vital for diverse studies of large populations aimed at evaluating the role of genetic factors in susceptibility to disease.
  
  
  OBJECTIVE
  This study was to test a protocol for the extraction of high quality genomic DNA from saliva samples obtained with mouthwash and taken from patients with periodontal disease.
  
  
  METHODS
  Saliva samples were taken from 60 patients and then stored at room temperature. DNA extraction was carried out at distinct post-sampling times (10, 20 and 30 days). Evaluation of genomic DNA was performed with spectrophotometry, electrophoresis, and PCR genotyping and sequencing.
  
  
  RESULTS
  The greatest concentration of DNA obtained was 352 μg at 10 days post-sampling, followed by 121.025 μg and 19.59 μg at 20 and 30 days, respectively. When determining the purity of DNA with the spectrophotometric ratio of 260/230, the relations of 1.20, 1.40 and 0.781 were obtained for 10, 20 and 30 days, respectively. In all samples, it was possible to amplify the product of 485 bp and the sequence of the amplicons showed 95% similarity to the reference sequence.
  
  
  CONCLUSIONS
  The present protocol represents an easy, safe and economical technique for obtaining high quality genomic DNA.
authors.affiliation: Posgrado en Investigación en Medicina, Laboratorio de Señalización Celular, Escuela Superior de Medicina, Instituto Politécnico Nacional, México, D.F., Mexico; Centro Interdisciplinario de Ciencias de la Salud, Unidad Santo Tomás, Instituto Politécnico Nacional, Av. de Los Maestros S/N, Col. Santo Tomas, Delegación Miguel Hidalgo, C.P. 11340 Ciudad de México, D.F., Mexico; Hospital General "Dr. Fernando Quiroz Gutiérrez", ISSSTE, General Felipe Ángeles y Canario S/N, Col. Bellavista, Delegación Álvaro Obregón, C.P. 01140 Ciudad de México, D.F., Mexico.
authors.forename: Ángel Chávez
authors.initials: ÁC
authors.lastname: Mendoza
authors.affiliation: Departamento de Genética, Unidad de Investigación, Instituto de Oftalmología "Conde de Valenciana", Chimalpopoca 14, Col. Obrera, Delegación Cuauhtémoc, C.P. 06800 Ciudad de México, D.F., Mexico.
authors.forename: Beatriz Buentello
authors.initials: BB
authors.lastname: Volante
authors.affiliation: Sección de Estudios de Posgrado e Investigación, Laboratorio de Señalización Celular, Escuela Superior de Medicina, Instituto Politécnico Nacional, Plan de San Luis y Díaz Mirón s/n, Col. Santo Tomas, Delegación Miguel Hidalgo, C.P. 11340 Ciudad de México, D.F., Mexico.
authors.forename: María Esther Ocharán
authors.initials: ME
authors.lastname: Hernández
authors.affiliation: Sección de Estudios de Posgrado e Investigación, Laboratorio de Señalización Celular, Escuela Superior de Medicina, Instituto Politécnico Nacional, Plan de San Luis y Díaz Mirón s/n, Col. Santo Tomas, Delegación Miguel Hidalgo, C.P. 11340 Ciudad de México, D.F., Mexico.
authors.forename: Claudia Camelia Calzada
authors.initials: CC
authors.lastname: Mendoza
authors.affiliation: Departamento de Inmunobioquimica, Instituto Nacional de Perinatología, Montes Urales 800 Col. Virreyes, Delegación Miguel Hidalgo, C.P. 11000 Ciudad de México, D.F., Mexico.
authors.forename: Arturo Flores
authors.initials: AF
authors.lastname: Pliego
authors.affiliation: Coordinación de Hematología Perinatal, Instituto Nacional de Perinatología, Montes Urales 800 Col. Virreyes, Delegación Miguel Hidalgo, C.P. 11000 Ciudad de México, D.F., Mexico.
authors.forename: Héctor A
authors.initials: HA
authors.lastname: Baptista Gonzalez
authors.affiliation: Coordinación de Hematología Perinatal, Instituto Nacional de Perinatología, Montes Urales 800 Col. Virreyes, Delegación Miguel Hidalgo, C.P. 11000 Ciudad de México, D.F., Mexico.
authors.forename: Higinio Estrada
authors.initials: HE
authors.lastname: Juárez
keywords: Chronic periodontitis; Genomic DNA; Mouthwash; PCR; Saliva