Characterizing multiple exogenous and endogenous small RNA populations in parallel with subfemtomolar sensitivity using a streptavidin gel-shift assay.

@article{Ebhardt2009CharacterizingME,
  title={Characterizing multiple exogenous and endogenous small RNA populations in parallel with subfemtomolar sensitivity using a streptavidin gel-shift assay.},
  author={H. Alexander Ebhardt and Peter J. Unrau},
  journal={RNA},
  year={2009},
  volume={15 4},
  pages={
          724-31
        }
}
Here we present a simple and inexpensive gel-shift assay for the detection and quantification of small RNAs. The assay is at least 5-10 times more sensitive than a conventional Northern, and is highly scalable. Total RNA is first size purified to enrich the desired size range, phosphatase treated, and then radiolabeled to high specific activity using polynucleotide kinase. The resulting RNA stock is then hybridized to an excess of biotinylated DNA probe oligonucleotide, prior to mixing with… CONTINUE READING

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