Issue 8, 2007
From the journal:

Molecular BioSystems

A novel SNP detection technique utilizing a multiple primer extension (MPEX) on a phospholipid polymer-coated surface

Kazuhide Imai,a   Yasukazu Ogai,b   Daisuke Nishizawa,b   Shinya Kasai,b   Kazutaka Ikedab  and  Hisashi Koga*a  

* Corresponding authors

a Laboratory of Medical Genomics, Department of Human Genome Technology, Kazusa DNA Research Institute, 2-6-7 Kazusa-Kamatari, Kisarazu, Chiba 292-0818, Japan
E-mail: hkoga@kazusa.or.jp
Fax: +81 438 52 3914
Tel: +81 438 52 3505

b Division of Psychobiology, Tokyo Institute of Psychiatry, 2-1-8 Kamikitazawa, Setagaya-ku, Tokyo 156-8585, Japan

Abstract

Conventional methods for detecting single nucleotide polymorphisms (SNPs), including direct DNA sequencing, pyrosequencing, and melting curve analysis, are to a great extent limited by their requirement for particular detection instruments. To overcome this limitation, we established a novel SNP detection technique utilizing multiple primer extension (MPEX) on a phospholipid polymer-coated surface. This technique is based on the development of a new plastic S-BIO® PrimeSurface® with a biocompatible polymer; its surface chemistry offers extraordinarily stable thermal properties, as well as chemical properties advantageous for enzymatic reactions on the surface. To visualize allele-specific PCR products on the surface, biotin-dUTP was incorporated into newly synthesized PCR products during the extension reaction. The products were ultimately detected by carrying out a colorimetric reaction with substrate solution containing 4-nitro-blue tetrazolium chloride (NBT) and 5-bromo-4-chloro-3-indolyl phosphate (BCIP). We demonstrated the significance of this novel SNP detection technique by analyzing representative SNPs on 4 LD blocks of the µ opioid receptor gene. We immobilized 20 allele-specific oligonucleotides on this substrate, and substantially reproduced the results previously obtained by other methods.

Graphical abstract: A novel SNP detection technique utilizing a multiple primer extension (MPEX) on a phospholipid polymer-coated surface

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Article information

DOI
https://doi.org/10.1039/B701645J
Article type
Method
Submitted
02 Feb 2007
Accepted
14 May 2007
First published
01 Jun 2007

Mol. BioSyst., 2007,3, 547-553

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A novel SNP detection technique utilizing a multiple primer extension (MPEX) on a phospholipid polymer-coated surface

K. Imai, Y. Ogai, D. Nishizawa, S. Kasai, K. Ikeda and H. Koga, Mol. BioSyst., 2007, 3, 547 DOI: 10.1039/B701645J

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