Single molecule FRET characterization of large ribozyme folding.
- Cardo L Institute of Inorganic Chemistry, University of Zurich, Zurich, Switzerland.
- Karunatilaka KS
- Rueda D
- Sigel RK
- 2012-02-09
Published in:
- Methods in molecular biology (Clifton, N.J.). - 2012
Base Sequence
Calibration
DEAD-box RNA Helicases
Fluorescence Polarization
Fluorescence Resonance Energy Transfer
Introns
Magnesium Chloride
Microfluidic Analytical Techniques
Molecular Sequence Data
Nucleic Acid Conformation
Polyethylene Glycols
RNA, Catalytic
Saccharomyces cerevisiae
Saccharomyces cerevisiae Proteins
Transcription, Genetic
English
A procedure to investigate the folding of group II intron by single molecule Fluorescence Resonance Energy Transfer (smFRET) using total internal reflection fluorescence microscopy (TIRFM) is described in this chapter. Using our previous studies on the folding and dynamics of a large ribozyme in the presence of metal ions (i.e., Mg(2+) and Ca(2+)) and/or the DEAD-box protein Mss116 as an example, we here describe step-by-step procedures to perform experiments. smFRET allows the investigation of individual molecules, thus, providing kinetic and mechanistic information hidden in ensemble averaged experiments.
- Language
-
- English
- Open access status
- green
- Identifiers
-
- DOI 10.1007/978-1-61779-545-9_15
- PMID 22315073
- Persistent URL
- https://sonar.ch/global/documents/65865
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