@inbook{5140a9637d334540b101ede1bb912310,
title = "Analysis of duplex unwinding by RNA helicases using stopped-flow fluorescence spectroscopy",
abstract = "The characterization of unwinding reactions by RNA helicases often requires the determination of rate constants that are too fast to be measured by traditional, manual gel-based methods. Stopped-flow fluorescence measurements allow access to fast unwinding rate constants. In this chapter, we outline strategies and experimental considerations for the design of stopped-flow fluorescence experiments to monitor duplex unwinding by RNA helicases, with focus on DEAD-box helicases. We discuss advantages, disadvantages, and technical considerations for stopped-flow approaches, as well as substrate design. In addition, we list protocols and explain functional information obtained with these experiments.",
keywords = "ATP, Base pair, Fluorescence, Helix, Kinetics, RNA, Stopped flow",
author = "Andrea Putnam and Eckhard Jankowsky",
note = "Funding Information: We thank the members of our laboratory for many fruitful discussions. We are grateful to Dr. Michael Harris (Case Western Reserve University) for the opportunity to use the stopped-flow fluorimeter and to Dr. Piet deBoer (Case Western Reserve University) for the opportunity to use the steady-state fluorimeter. Research on RNA helicases in our laboratory is supported by the NIH (GM 067700 to E. J.).",
year = "2012",
doi = "10.1016/B978-0-12-396546-2.00001-2",
language = "English (US)",
series = "Methods in Enzymology",
publisher = "Academic Press Inc.",
pages = "1--27",
booktitle = "Methods in Enzymology",
}