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.
Sensors 2017, 17(6), 1201; https://doi.org/10.3390/s17061201
Received: 13 March 2017 / Revised: 17 May 2017 / Accepted: 18 May 2017 / Published: 24 May 2017
(This article belongs to the Special Issue Genosensing)
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
▼
Show Figures
Figure 1
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited
- Supplementary File 1:
PDF-Document (PDF, 754 KiB)
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. https://doi.org/10.3390/s17061201
AMA Style
Fjelstrup S, Andersen MB, Thomsen J, Wang J, Stougaard M, Pedersen FS, Ho Y-P, Hede MS, Knudsen BR. The Effects of Dithiothreitol on DNA. Sensors. 2017; 17(6):1201. https://doi.org/10.3390/s17061201
Chicago/Turabian StyleFjelstrup, Søren, Marie B. Andersen, Jonas Thomsen, Jing Wang, Magnus Stougaard, Finn S. Pedersen, Yi-Ping Ho, Marianne S. Hede, and Birgitta R. Knudsen 2017. "The Effects of Dithiothreitol on DNA" Sensors 17, no. 6: 1201. https://doi.org/10.3390/s17061201
Find Other Styles
Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.
