Fluorescence-based on-line detection as an analytical tool in RNA electrophoresis.
In this chapter, we present methods for denaturing and native gel electrophoresis of nucleic acids based on fluorescently labeled probes and automatic signal detection by a deoxyribonucleic acid sequencer. Specific examples are given for the determination of ribonucleic acid (RNA) fragmentation patterns, of (deoxy)ribozyme kinetics, and of direct or competitive gel-shift assays. These methods can replace widely used radioisotope-based protocols, for example, for secondary structure mapping of RNA and for the characterization of nucleic acid ligand interactions.
Scheffer
Ute
U
Department of Organic Chemistry and Cheical Biology, Johann Wolfgang Goethe-University Frankfurt, Franfurt am Main, Germany.
Göbel
Michael
M
D001483:Base Sequence; D004586:Electrophoresis; D007700:Kinetics; D012313:RNA / Q000737:chemistry / Q000302:isolation & purification*; D013050:Spectrometry, Fluorescence / Q000379:methods*
D012313:RNA