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Electrochemical detection of β-1,3-glucanase gene from transgenic capsicums using asymmetric PCR generated by a detecting probe and an anchoring probe

Paper ID Volume ID Publish Year Pages File Format Full-Text
24565 43527 2010 8 PDF Available
Title
Electrochemical detection of β-1,3-glucanase gene from transgenic capsicums using asymmetric PCR generated by a detecting probe and an anchoring probe
Abstract

A design for recognition of β-1,3-glucanase gene (Glu) specific sequence based on probe extension was described. The detecting probe DNA and the anchoring prober were hybridized with the same target DNA firstly, then the probes were extended by DNA polymerase reaction. After that the double strand DNA was denatured, and the extended detecting probe was immobilized on a glassy carbon electrode via nanoparticle gold (AuNP). In electrochemical detection (cyclic voltammetry, CV and differential pulse voltammetry, DPV), an increased peak current (ip) of the indicator (methylene blue, MB) was obtained compared with the probe without extension. Three differently long DNAs of Glu specific sequence were employed as the target: oligonucleotide acid, molecular cloning vector DNA and total genome DNA of transgenic capsicum. The estimated DPV detection limits for three targets of oligonucleotide, the molecular cloning vector DNA and genome DNA were 2.6 × 10−13, 6.0 × 10−13 and 8.0 × 10−13 mol l−1 respectively.

Keywords
β-1,3-Glucanase gene; Transgenic capsicum; DNA detection; Asymmetric polymerase chain reaction; Electrochemical assay
First Page Preview
Electrochemical detection of β-1,3-glucanase gene from transgenic capsicums using asymmetric PCR generated by a detecting probe and an anchoring probe
Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 145, Issue 1, 1 January 2010, Pages 1–8
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering