title: DNA amplification using phi29 DNA polymerase validates gene polymorphism analysis from buccal mucosa samples.
abstract: Venous blood is currently the most common source of DNA for gene polymorphism screening; however, blood sampling is invasive and difficult to perform in general dental treatment. Buccal mucosa samples provide an alternative source of DNA, but it is frequently difficult to effectively amplify the DNA owing to the small amounts of sample material obtained. This study was performed to establish a method for performing total genomic DNA amplification from buccal mucosa samples using phi29 DNA polymerase. Total genomic DNA was isolated from buccal mucosa samples obtained from healthy subjects and was amplified using phi29 DNA polymerase. To determine the suitability of the extracted DNA for genotyping, polymerase chain reaction and restriction fragment length polymorphism analyses were performed for the IL-1 gene polymorphism. Genotyping of the IL-1 polymorphism was successful using the amplified DNA from a buccal mucosa, but genotyping was unsuccessful using the unamplified control because of low DNA purity. The method of extracting DNA from a buccal mucosa is painless, simple, minimally invasive, and rapid. Genomic DNA from a buccal mucosa can be amplified by phi29 DNA polymerase in sufficient quantity and quality to conduct gene polymorphism analyses.
authors.affiliation: Department of Oral Reconstruction and Rehabilitation, Kyushu Dental College, Graduate School, Japan.
authors.forename: Ryoji
authors.initials: R
authors.lastname: Taniguchi
authors.forename: R
authors.initials: R
authors.lastname: Taniguchi
authors.forename: Chihiro
authors.initials: C
authors.lastname: Masaki
authors.forename: C
authors.initials: C
authors.lastname: Masaki
authors.forename: Yuhi
authors.initials: Y
authors.lastname: Murashima
authors.forename: Y
authors.initials: Y
authors.lastname: Murashima
authors.forename: Michiko
authors.initials: M
authors.lastname: Makino
authors.forename: M
authors.initials: M
authors.lastname: Makino
authors.forename: Tatsuro
authors.initials: T
authors.lastname: Kojo
authors.forename: T
authors.initials: T
authors.lastname: Kojo
authors.forename: Tetsuji
authors.initials: T
authors.lastname: Nakamoto
authors.forename: T
authors.initials: T
authors.lastname: Nakamoto
authors.forename: Ryuji
authors.initials: R
authors.lastname: Hosokawa
authors.forename: R
authors.initials: R
authors.lastname: Hosokawa
mesh_terms: D004259:DNA-Directed DNA Polymerase / Q000032:analysis; D006801:Humans; D007375:Interleukin-1 / Q000235:genetics; D009061:Mouth Mucosa / Q000737:chemistry*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D011110:Polymorphism, Genetic; D014764:Viral Proteins
chemical_list: D007375:Interleukin-1; D014764:Viral Proteins; C060516:phage phi29 protein p1; D004259:DNA-Directed DNA Polymerase