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Southern blotting

Abstract

This protocol describes a basic method to perform the Southern blot. Blotting allows the detection of specific molecules among a mixture separated by gel electrophoresis. Molecules are transferred from the gel to a porous membrane by capillary action using absorbent paper to soak solution through the gel and the membrane. For DNA, specific sequences are detected in the membrane by molecular hybridization with labeled nucleic acid probes. The original method, on which this protocol is based, used labeled RNAs to detect specific DNA fragments in genomic DNA that had been digested with restriction endonucleases. This protocol can be completed in 1–5 d and is inexpensive to carry out, as it requires only basic laboratory equipment.

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Figure 1: Steps in the procedure for transferring DNA from an agarose gel strip to a hybridization membrane.
Figure 2: Transfer from wide gels.
Figure 3: Example of a Southern blot.

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Acknowledgements

Where possible, a reference is given to credit the original authors or inventors; in many cases there is likely to be a more recent publication which would be more useful or informative, but it was not felt possible or appropriate to include all of these references in this short article.

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Correspondence to Ed Southern.

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Southern, E. Southern blotting. Nat Protoc 1, 518–525 (2006). https://doi.org/10.1038/nprot.2006.73

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