real-time assay for monitoring nucleic acid cleavage by quadruplex formation

Direct and straightforward methods to follow nucleic acid cleavage are needed. A spectrophotometric quadruplex formation assay (QFA) was developed, which allows real-time monitoring of site-specific cleavage of nucleic acids. QFA was applied to study both protein and nucleic acid restriction enzymes...

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Bibliographic Details
Published in:Nucleic acids research 2006-11, Vol.34 (20), p.e141-e141
Main Author: Kankia, Besik I
Format: Article
Language:English
Subjects:
Aptamers, Nucleotide - analysis
Circular Dichroism
Deoxyribonuclease EcoRI - metabolism
DNA - analysis
DNA - metabolism
DNA Restriction Enzymes - metabolism
DNA, Catalytic - metabolism
G-Quadruplexes
Kinetics
Methods Online
RNA - metabolism
Spectrophotometry, Ultraviolet - methods
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Summary:Direct and straightforward methods to follow nucleic acid cleavage are needed. A spectrophotometric quadruplex formation assay (QFA) was developed, which allows real-time monitoring of site-specific cleavage of nucleic acids. QFA was applied to study both protein and nucleic acid restriction enzymes, and was demonstrated to accurately determine Michaelis-Menten parameters for the cleavage reaction catalyzed by EcoRI. QFA can be used to study the mechanisms of protein-nucleic acid recognition. QFA is also a useful tool for dissecting individual nicking rates of a double-stranded cleavage.
ISSN:0305-1048
1362-4962
DOI:10.1093/nar/gkl594