Journal
BIOSENSORS & BIOELECTRONICS
Volume 41, Issue -, Pages 526-531Publisher
ELSEVIER ADVANCED TECHNOLOGY
DOI: 10.1016/j.bios.2012.09.017
Keywords
Electrochemical DNA (E-DNA) sensor; Surface initiated enzymatic polymerization (SIEP); Terminal deoxynucleotidyl transferase (TdT); TdT-mediated extension reaction
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Funding
- National Science Foundation [21105048, 21104030]
- NUST [2011XQTR08]
- Promotional Foundation for the Excellent Middle-Aged and Young Scientists of Shandong Province of China [BS2009SW040]
- Project of Shandong Province Higher Educational Science and Technology Program [J11LB03]
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In this work, we report a novel strategy of electrochemical DNA (E-DNA) sensor based on surface initiated enzymatic polymerization (SIEP). This DNA sensor employs a capture DNA probe labeled with thiol at its 3' terminal to be immobilized at gold electrode via gold-thiol chemistry. Oligo (ethylene glycol) -terminated thiols (SH-OEGs) are then used to prepare an oligonucleotide-incorporated nonfouling surface (ONS). After the sequence-specific recognition of target DNA, terminal deoxynucleotidyl transferase (TdT) is employed to catalyze the sequential addition of deoxynucleotides (dNTPs) at the 3'-OH group of target DNA without template. During the TdT-mediated extension reaction, by using biotinlated 2'-deoxyadenosine 5'-triphosphate (biotin-dATP), biotin labels are incorporated into the SIEP-generated long single-stranded DNA (ssDNA). Specific binding of avidin-horseradish peroxidase (Av-HRP) to the biotin label leads to enzyme turnover-based signal transduction. By using this new strategy, we demonstrated the high picomolar sensitivity and a broad detection range of six orders of magnitude. (C) 2012 Elsevier B.V. All rights reserved.
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