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research article

Single Molecule Localization and Discrimination of DNA–Protein Complexes by Controlled Translocation Through Nanocapillaries

Bulushev, Roman D.  
•
Marion, Sanjin  
•
Petrova, Ekaterina  
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2016
Nano Letters

Through the use of optical tweezers we performed controlled translocations of DNA-protein complexes through nanocapillaries. We used RNA polymerase (RNAP) with two binding sites on a 7.2 kbp DNA fragment and a dCas9 protein tailored to have five binding sites on lambda-DNA (48.5 kbp). Measured localization of binding sites showed a shift from the expected positions on the DNA that we explained using both analytical fitting and a stochastic model. From the measured force versus stage curves we extracted the non equilibrium work done during the translocation of a DNA-protein complex and used it to obtain an estimate of the effective charge of the complex. In combination with conductivity measurements, we provided a proof of concept for discrimination between different DNA protein complexes simultaneous to the localization of their binding sites.

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Type
research article
DOI
10.1021/acs.nanolett.6b04165
Web of Science ID

WOS:000389963200084

Author(s)
Bulushev, Roman D.  
Marion, Sanjin  
Petrova, Ekaterina  
Davis, Sebastian J.  
Maerkl, Sebastian J.  
Radenovic, Aleksandra  
Date Issued

2016

Publisher

American Chemical Society (ACS)

Published in
Nano Letters
Volume

16

Issue

12

Start page

7882

End page

7890

Subjects

Nanopore

•

nanocapillary

•

force measurements

•

DNA-protein complex

•

protein binding site

•

Jarzynski equation

Editorial or Peer reviewed

REVIEWED

Written at

EPFL

EPFL units
LBEN  
LBNC  
Available on Infoscience
December 7, 2016
Use this identifier to reference this record
https://infoscience.epfl.ch/handle/20.500.14299/131841
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