NDLI: Sandwich structure electrochemical assay for single stranded DNA detection

Content Provider	IEEE Xplore Digital Library
Author	I-Jane Chen White, I.M.
Copyright Year	2010
Description	Author affiliation: Fischell Department of Bioengineering, University of Maryland, College Park, USA (I-Jane Chen; White, I.M.)
Abstract	A novel DNA sequence biosensor that utilizes enzyme-labeled probes and electrochemical detection via an interdigitated array (IDA) electrode on a microfluidic chip is presented. Sandwich structure DNA sequence recognition is performed by immobilizing a single stranded DNA (ssDNA) on the microchannel surface (primary probe) that is complementary to a portion of the target, followed by hybridization with target ssDNA and a second ssDNA (secondary DNA probe), which is complementary to another region of the target and is conjugated with β-Galactosidase (β-Gal). Surface confined β-Gal is detected by introducing its substrate, 4-aminophenyl-D-galactopyranoside (PAPG), into the system; β-Gal cleaves PAPG into the electrochemical active compound, p-amino phenol (PAP), which is detected by a Au IDA in a microfluidic channel. The signal is amplified by cycling its reduced and oxidized form between the interleaved anode and cathode. On the chips which are integrated in 60µm high channels, the oxidation currents show linear correlation to the immobilized target DNA concentration between 0 and 100nM. Currents collected on chips where 0 and 0.5nM target DNA is immobilized are significantly different. Similarly, on the chips integrated in the 35µm channels where target DNA is not introduced to form the sandwich (negative control), the oxidation current shows non-differentiable results whether PAPG is present or absent in buffer solution. On the contrary, on the chip where 0.1nM target DNA hybridized with probes, the oxidation current is significantly higher when the buffer contains PAPG. As the enzymatic reaction time increases from 15 to 30 seconds, the signal difference between PAPG and buffer solution almost doubled. By reducing the channel height from 60 to 35µm, the total assay time is reduced from 80 minutes to 35 minutes.
Starting Page	470
Ending Page	473
File Size	609675
Page Count	4
File Format	PDF
ISBN	9781424481705
ISSN	19300395
e-ISBN	9781424481699
e-ISBN	9781424481682
DOI	10.1109/ICSENS.2010.5690168
Language	English
Publisher	Institute of Electrical and Electronics Engineers, Inc. (IEEE)
Publisher Date	2010-11-01
Publisher Place	USA
Access Restriction	Subscribed
Rights Holder	Institute of Electrical and Electronics Engineers, Inc. (IEEE)
Subject Keyword	DNA Probes Microchannel Oxidation Electrodes Sandwich structures Microfluidics
Content Type	Text
Resource Type	Article
Subject	Electrical and Electronic Engineering

Sl.	Authority	Responsibilities	Communication Details
1	Ministry of Education (GoI), Department of Higher Education	Sanctioning Authority	https://www.education.gov.in/ict-initiatives
2	Indian Institute of Technology Kharagpur	Host Institute of the Project: The host institute of the project is responsible for providing infrastructure support and hosting the project	https://www.iitkgp.ac.in
3	National Digital Library of India Office, Indian Institute of Technology Kharagpur	The administrative and infrastructural headquarters of the project	Dr. B. Sutradhar bsutra@ndl.gov.in
4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
5	Website/Portal (Helpdesk)	Queries regarding NDLI and its services	support@ndl.gov.in
6	Contents and Copyright Issues	Queries related to content curation and copyright issues	content@ndl.gov.in
7	National Digital Library of India Club (NDLI Club)	Queries related to NDLI Club formation, support, user awareness program, seminar/symposium, collaboration, social media, promotion, and outreach	clubsupport@ndl.gov.in
8	Digital Preservation Centre (DPC)	Assistance with digitizing and archiving copyright-free printed books	dpc@ndl.gov.in
9	IDR Setup or Support	Queries related to establishment and support of Institutional Digital Repository (IDR) and IDR workshops	idr@ndl.gov.in

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