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Northern blotting: transfer of denatured RNA to membranes

Northern blotting and hybridization are used to study gene expression by detecting RNA species of interest, and to identify alternate RNA splicing patterns. It is analogous to Southern blotting, which is used to analyze DNA. This protocol describes the transfer of RNA from agarose gels to nylon membranes and the fixation of the RNA to the membrane1,2,3.

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References

  1. Alwine, J.C., Kemp, D.J., & Stark, G.R. Method for detection of specific RNAs in agarose gels by transfer to diazobenzyloxymethyl-paper and hybridization with DNA probes. Proc. Natl. Acad. Sci. USA 74, 5350–5354 (1977).

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  2. Angeletti, B., Battiloro, E., Pascale, E., & D'Ambrosio, E. Southern and northern blot fixing by microwave oven. Nucleic Acids Res. 23, 879–880 (1995).

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  3. Herrin, D.L. & Schmidt, G.W. Rapid, reversible staining of northern blots prior to hybridization. BioTechniques 6, 196–197, 199–200 (1988).

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Northern blotting: transfer of denatured RNA to membranes. Nat Methods 2, 997–998 (2005). https://doi.org/10.1038/nmeth1205-997

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  • DOI: https://doi.org/10.1038/nmeth1205-997

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