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Sensors 2017, 17(6), 1201; doi:10.3390/s17061201

The Effects of Dithiothreitol on DNA

1
Department of Molecular Biology and Genetics, Aarhus University, 8000 Aarhus, Denmark
2
Interdisciplinary Nanoscience Center (iNANO), Aarhus University, 8000 Aarhus, Denmark
3
Department of Pathology, Aarhus University Hospital, 8000 Aarhus, Denmark
4
Division of Biomedical Engineering, Department of Electronic Engineering, The Chinese University of Hong Kong, Shatin, NT, Hong Kong SAR, China
5
Zymonostics ApS, 8000 Aarhus, Denmark
*
Authors to whom correspondence should be addressed.
Received: 13 March 2017 / Revised: 17 May 2017 / Accepted: 18 May 2017 / Published: 24 May 2017
(This article belongs to the Special Issue Genosensing)
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Abstract

With the novel possibilities for detecting molecules of interest with extreme sensitivity also comes the risk of encountering hitherto negligible sources of error. In life science, such sources of error might be the broad variety of additives such as dithiothreitol (DTT) used to preserve enzyme stability during in vitro reactions. Using two different assays that can sense strand interruptions in double stranded DNA, we here show that DTT is able to introduce nicks in the DNA backbone. DTT was furthermore shown to facilitate the immobilization of fluorescent DNA on an NHS-ester functionalized glass surface. Such reactions may in particular impact the readout from single molecule detection studies and other ultrasensitive assays. This was highlighted by the finding that DTT markedly decreased the signal to noise ratio in a DNA sensor based assay with single molecule resolution. View Full-Text
Keywords: single molecule detection; DTT; DNA modifying enzyme; DNA sensor; thiol; DNA nicking single molecule detection; DTT; DNA modifying enzyme; DNA sensor; thiol; DNA nicking
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MDPI and ACS Style

Fjelstrup, S.; Andersen, M.B.; Thomsen, J.; Wang, J.; Stougaard, M.; Pedersen, F.S.; Ho, Y.-P.; Hede, M.S.; Knudsen, B.R. The Effects of Dithiothreitol on DNA. Sensors 2017, 17, 1201.

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