Authors: Maekawa M; Taniguchi T; Uramoto T; Higashi H; Horii T; Takeshita A; Sugimura H; Kanamori M
Abstract: BACKGROUND: To investigate relationships between phenotypes and genotypes is not simple. We propose a phenotype-to-genotype screening strategy and pooled DNA system. As a pilot study of this strategy, we used arbitrarily primed polymerase chain reaction (AP-PCR) in combination with single-stranded DNA conformation polymorphism (SSCP) to screen for genetic polymorphisms associated with longevity. METHODS: Study subjects were separated into 3 age groups, individuals aged >100 years, 90-99 years and 60-69 years. Genomic DNAs were prepared from each individual, pooled to represent the 5 study groups, and then the pooled genomic DNAs were subjected to AP-PCR-SSCP analysis. RESULTS: We found 1 SNP more frequently in senior citizens with longevity. The genotype frequency of the 82133G>A polymorphism of human chromosome 3 clone RP11-61K12 (AC011199) differed significantly (P=0.0189, Fisher's exact test) between older subjects (>90 years) and younger subjects (<70 years). It is noteworthy that the strategy we describe herein was useful for identifying an SNP that showed statistically significant differences in its distribution across the subject groups. CONCLUSIONS: The pooled DNA strategy and quantitative genotype discrimination can also be applied to screening for the relationship between phenotype and genotype more effectively.Keywords: Adult; Aged; Aged, 80 and over; DNA, Single-Stranded/chemistry/*genetics; Genotype; Humans; Longevity/*genetics; Middle Aged; Nucleic Acid Conformation; Phenotype; Pilot Projects; Polymerase Chain Reaction/*methods; *Polymorphism, Genetic
Journal: Clinica chimica acta; international journal of clinical chemistry
Date: April 12, 2005
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Maekawa M, Taniguchi T, Uramoto T, Higashi H, Horii T, Takeshita A, Sugimura H, Kanamori M (2005) Pilot study of arbitrarily primed PCR-single stranded DNA conformation polymorphism analysis for screening genetic polymorphisms related to specific phenotypes. Clinica chimica acta; international journal of clinical chemistry 355: 181-4.