NDLI: SNP Genotyping by Gel-immobilized ssDNA and Biolumometric Assay Coupled with Allele-specific Primer Extension Reaction

Content Provider	IEEE Xplore Digital Library
Author	Pengfeng Xiao Huan Huang Haipng Wu Xiaodang Zhang Guohua Zhou
Copyright Year	2006
Description	Author affiliation: Key Lab. of Bioelectronics, Southeast Univ., Nanjing (Pengfeng Xiao)
Abstract	The new gel-immobilization method for preparing a single stranded DNA (ssDNA) template has been described for single nucleotide polymorphisms (SNPs) genotyping by bioluminometric assay based on allele-specfic extension reaction. In this method, PCR products amplified by a primer modified with acrylamide group at the 5'-terminal is copolymerized with acrylamide monomer to form gel. After the ssDNA immobilized on gel being denatured, the impurities are removed by electrophoresis. SNPs genotyping is performed by allele-specific primer extension reaction. The SNP type was identified by comparing the signal intensities from the extension by two allele specific primers hybridized on gel-mobilized ssDNA. If the 3' end of a primer is complementary to the template, polymerase extension occurs and pyrophosphate (PPi) released. The signal is produced by using PPi conversion and luciferase reactions. As many bases are extended in one extension reaction, the sensitivity of the method is very high. This new strategy of the ssDNA immobilization is robust, simple, and inexpensive. It can be used for various application of DNA analysis, such as SNP genotyping and pyrosequencing.
Starting Page	734
Ending Page	738
File Size	4565791
Page Count	5
File Format	PDF
ISBN	1424401399
DOI	10.1109/NEMS.2006.334884
Language	English
Publisher	Institute of Electrical and Electronics Engineers, Inc. (IEEE)
Publisher Date	2006-01-18
Publisher Place	China
Access Restriction	Subscribed
Rights Holder	Institute of Electrical and Electronics Engineers, Inc. (IEEE)
Subject Keyword	DNA Polymers Electrokinetics Sequences Robustness Surface contamination Purification Chemistry Glass Systems engineering and theory single-nucleotide polymorphism allele-specific extension acrylamide-modified nucleic acids gel-immobilization
Content Type	Text
Resource Type	Article

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