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

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Contents

Hot off the pressHot off the RSC press ( Molecular BioSystems Vol - 3 Issue - 8 )

Proteomics data validation: why all must provide data

Deduction of intracellular sub-systems from a topological description of the network

Transgenic approaches to study wing color pattern development in Lepidoptera

Reverse interactomics: decoding protein–protein interactions with combinatorial peptide libraries

Dynamic proteomics in mammalian cells: capabilities and challenges

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

Identification of Conus amadis disulfide isomerase: minimum sequence length of peptide fragments necessary for protein annotation

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

Content Provider	Royal Society of Chemistry (RSC)
Author	Nishizawa, Daisuke Ikeda, Kazutaka Imai, Kazuhide Ogai, Yasukazu Kasai, Shinya Koga, Hisashi
Copyright Year	2007
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.
Starting Page	547
Ending Page	553
Page Count	7
File Format	HTM / HTML PDF
ISSN	1742206X
Volume Number	3
Issue Number	8
Journal	Molecular BioSystems
DOI	10.1039/b701645j
Language	English
Publisher	Royal Society of Chemistry
Access Restriction	Open
Subject Keyword	NBT 5-Bromo-4-chloro-3-indolyl phosphate Opioid receptor Phospholipid Melting curve analysis Polymer Single-nucleotide polymorphism Tetrazolium chloride Nitro blue tetrazolium chloride Polymerase chain reaction Phosphate Pyrosequencing Biocompatibility Surface science DNA sequencing
Content Type	Text
Resource Type	Article
Subject	Molecular Biology Biotechnology

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In previous studies, proliferating chondrocytes were identified on the edge of the laser-irradiated regions in cartilage tissue using flow cytometry and whole mount double antibody system (Nitro blue tetrazolium 5- bromo-4-chloro-3-indolyl phosphate NBT/BCIP antibody staining) [1]. For these studies

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

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