Imperial College London
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Professor David S. Rueda

Faculty of Medicine, Department of Infectious Disease

Chair in Molecular and Cellular Biophysics
 
 
 
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Contact

 

david.rueda Website

 
 
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Location

 

6.12DLMS BuildingHammersmith Campus

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Summary

 

Publications

Citation

BibTex format

@article{Belan:2021:10.1016/j.xpro.2021.100588,
author = {Belan, O and Moore, G and Kaczmarczyk, A and Newton, MD and Anand, R and Boulton, SJ and Rueda, DS},
doi = {10.1016/j.xpro.2021.100588},
journal = {STAR Protocols},
pages = {1--18},
title = {Generation of versatile ss-dsDNA hybrid substrates for single-molecule analysis.},
url = {http://dx.doi.org/10.1016/j.xpro.2021.100588},
volume = {2},
year = {2021}
}
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RIS format (EndNote, RefMan)

TY  - JOUR
AB  - Here, we describe a rapid and versatile protocol to generate gapped DNA substrates for single-molecule (SM) analysis using optical tweezers via site-specific Cas9 nicking and force-induced melting. We provide examples of single-stranded (ss) DNA gaps of different length and position. We outline protocols to visualize these substrates by replication protein A-enhanced Green Fluorescent Protein (RPA-eGFP) and SYTOX Orange staining using commercially available optical tweezers (C-TRAP). Finally, we demonstrate the utility of these substrates for SM analysis of bidirectional growth of RAD-51-ssDNA filaments. For complete details on the use and execution of this protocol, please refer to Belan et al. (2021).
AU  - Belan,O
AU  - Moore,G
AU  - Kaczmarczyk,A
AU  - Newton,MD
AU  - Anand,R
AU  - Boulton,SJ
AU  - Rueda,DS
DO  - 10.1016/j.xpro.2021.100588
EP  - 18
PY  - 2021///
SN  - 2666-1667
SP  - 1
TI  - Generation of versatile ss-dsDNA hybrid substrates for single-molecule analysis.
T2  - STAR Protocols
UR  - http://dx.doi.org/10.1016/j.xpro.2021.100588
UR  - https://www.ncbi.nlm.nih.gov/pubmed/34169285
UR  - https://www.sciencedirect.com/science/article/pii/S2666166721002951?via%3Dihub
UR  - http://hdl.handle.net/10044/1/93360
VL  - 2
ER  -
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