Rapid and Cost-Effective Genotyping Protocol for Angiotensin-Converting Enzyme Insertion/Deletion (Ins/Del) Polymorphism from Saliva

Madoka Kisoi; Miwako Moritsugu; Miho Imai; Kae Fukumoto; Yui Sakaguchi; Shigenori Murata; Sayuri Kawai; Atsushi Ichikawa; Kenji Kinoshita

doi:10.1248/bpb.b19-00110

Regular Articles

Rapid and Cost-Effective Genotyping Protocol for Angiotensin-Converting Enzyme Insertion/Deletion (Ins/Del) Polymorphism from Saliva

Madoka Kisoi, Miwako Moritsugu, Miho Imai, Kae Fukumoto, Yui Sakaguchi, Shigenori Murata, Sayuri Kawai, Atsushi Ichikawa, Kenji Kinoshita

Author information

Madoka Kisoi
School of Pharmaceutical Sciences, Mukogawa Women’s University
Miwako Moritsugu
School of Pharmaceutical Sciences, Mukogawa Women’s University
Miho Imai
School of Pharmaceutical Sciences, Mukogawa Women’s University
Kae Fukumoto
School of Pharmaceutical Sciences, Mukogawa Women’s University
Yui Sakaguchi
School of Pharmaceutical Sciences, Mukogawa Women’s University
Shigenori Murata
School of Pharmaceutical Sciences, Mukogawa Women’s University
Institute of Biosciences, Mukogawa Women’s University
Sayuri Kawai
Institute of Biosciences, Mukogawa Women’s University
Atsushi Ichikawa
Institute of Biosciences, Mukogawa Women’s University
Kenji Kinoshita Corresponding author
School of Pharmaceutical Sciences, Mukogawa Women’s University
Institute of Biosciences, Mukogawa Women’s University

Keywords: angiotensin-converting enzyme (ACE) gene, melting curve analysis, real-time PCR, insertion/deletion (Ins/Del) polymorphism, water-soluble paper, oral mucosal cell

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2019 Volume 42 Issue 8 Pages 1345-1349

DOI https://doi.org/10.1248/bpb.b19-00110

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Abstract

DNA extraction and purification have been generally considered to be required for PCR assay. We demonstrated a new protocol using biological specimens directly as templates for real-time PCR with melting curve analysis. We confirmed the melting curve analysis was particularly suitable for the identification of the insertion/deletion (Ins/Del) polymorphism of the angiotensin-converting enzyme (ACE) gene. The new protocol we developed can be set up using simple and complete PCR analysis including data interpretation in under four hours with additional advantages of application for large-scale clinical research, diagnostics, and epidemiological studies at low cost.

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