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DNA Polymerase Activity Assay Using Near-infrared Fluorescent Labeled DNA Visualized by Acrylamide Gel Electrophoresis
Published on: October 6, 2017
A fluorescence biosensor based on double-stranded DNA and a cationic conjugated polymer coupled with exonuclease III
Yufei Liu1, Suyu Meng1, Jingjing Qin2
1School of Pharmacy, Xinxiang Medical University, Xinxiang, Henan 453003, PR China.
Abstract:
As a toxic substance on human health produced in food thermal treatment, simple analytical approaches are highly desired for the detection of acrylamide (ACR) in foods. With the aid of exonuclease III (Exo III), a simple fluorescence sensor was proposed based on carboxyfluorescein-labeled double-stranded DNA (FAM-dsDNA) and a cationic conjugated polymer (PFP). Fluorescence resonance energy transfer (FRET) efficiency between FAM and PFP was changed with and without ACR. When ACR was present, ACR and single-stranded DNA (P1, ssDNA) formed an adduct, allowing free FAM-labeled complementarity strand DNA (P2, FAM-csDNA) to appear in the solution and avoiding the digestion of P2 by Exo III. After the addition of PFP, the interaction of PFP and FAM induced strong FRET. Under optimized conditions, ACR was detected with a limit of detection (LOD) of 0.16 μM. According to this biosensor, a LOD of 1.3 μM in water extract samples was observed with a good recovery rate (95-110 %).

