@article{Robinson:2014:10.1021/ac502732s,
author = {Robinson, T and Valluri, P and Kennedy, G and Sardini, A and Dunsby, C and Neil, MAA and Baldwin, GS and French, PMW and de, Mello AJ},
doi = {10.1021/ac502732s},
journal = {Analytical Chemistry},
pages = {10732--10740},
title = {Analysis of DNA Binding and Nucleotide Flipping Kinetics Using Two-Color Two-Photon Fluorescence Lifetime Imaging Microscopy},
url = {http://dx.doi.org/10.1021/ac502732s},
volume = {86},
year = {2014}
}

Download

TY  - JOUR
AB  - Uracil DNA glycosylase plays a key role in DNA maintenance via base excision repair. Its role is to bind to DNA, locate unwanted uracil, and remove it using a base flipping mechanism. To date, kinetic analysis of this complex process has been achieved using stopped-flow analysis but, due to limitations in instrumental dead-times, discrimination of the “binding” and “base flipping” steps is compromised. Herein we present a novel approach for analyzing base flipping using a microfluidic mixer and two-color two-photon (2c2p) fluorescence lifetime imaging microscopy (FLIM). We demonstrate that 2c2p FLIM can simultaneously monitor binding and base flipping kinetics within the continuous flow microfluidic mixer, with results showing good agreement with computational fluid dynamics simulations.
AU  - Robinson,T
AU  - Valluri,P
AU  - Kennedy,G
AU  - Sardini,A
AU  - Dunsby,C
AU  - Neil,MAA
AU  - Baldwin,GS
AU  - French,PMW
AU  - de,Mello AJ
DO  - 10.1021/ac502732s
EP  - 10740
PY  - 2014///
SN  - 0003-2700
SP  - 10732
TI  - Analysis of DNA Binding and Nucleotide Flipping Kinetics Using Two-Color Two-Photon Fluorescence Lifetime Imaging Microscopy
T2  - Analytical Chemistry
UR  - http://dx.doi.org/10.1021/ac502732s
UR  - http://hdl.handle.net/10044/1/25097
VL  - 86
ER  - 

Download