Using FAM labeled DNA oligos to do RNA electrophoretic mobility shift assay.
The electrophoretic mobility shift assay is a useful tool to identify proteins and nucleic acids interactions. Traditionally, the nucleic acids fragments are end-labeled with (32)P. We present here the use of fluorescent methodologies for studies of RNA in place of radioactivity. The method is highly sensitive and quantitative with the use of an infrared fluorescence imaging system. Fluorescently labeled primers can be used to monitor protein-RNA interactions by fluorescent mobility shift assays. The simplicity and validity of this approach may have more advantages than that of previous methods that traditionally used hazardous radioisotopes. This method was successfully tested in the study of the interactions between MS2 Coat Protein and its RNA target.
Wang
Kun
K
Key Laboratory of MOE for Plant Developmental Biology, College of Life Sciences, Wuhan University, 430072 Wuhan, People's Republic of China.
Gao
Ya
Y
Peng
Xiaojue
X
Yang
Guohua
G
Gao
Feng
F
Li
Shaoqing
S
Zhu
Yingguo
Y
D036022:Capsid Proteins / Q000378:metabolism; D004247:DNA / Q000378:metabolism*; D024202:Electrophoretic Mobility Shift Assay / Q000379:methods*; D005452:Fluoresceins / Q000378:metabolism*; D005456:Fluorescent Dyes / Q000378:metabolism*; D017909:Levivirus / Q000378:metabolism; D058619:Maltose-Binding Proteins; D009841:Oligonucleotides; D033902:Periplasmic Binding Proteins / Q000378:metabolism; D012313:RNA / Q000378:metabolism*; D015203:Reproducibility of Results; D013194:Staining and Labeling
D036022:Capsid Proteins; D005452:Fluoresceins; D005456:Fluorescent Dyes; D058619:Maltose-Binding Proteins; D009841:Oligonucleotides; D033902:Periplasmic Binding Proteins; C024098:6-carboxyfluorescein; D012313:RNA; D004247:DNA