Southern Blotting and Related DNA Detection Techniques

Abstract

Southern blotting is a technique for transfer of DNA molecules from an electrophoresis gel to a nitrocellulose or nylon membrane, and is carried out prior to detection of specific molecules by hybridization probing.

Keywords: southern blotting; DNA preparation; DNA/DNA hybridization; hybridization probing; restriction fragment length polymorphism (RFLP)

Figure 1.

Southern blotting. Reproduced from Brown with permission of BIOS Scientific Publishers Ltd, Oxford.

Figure 2.

Detection of an RFLP by Southern blotting. In this example, lane 2 contains DNA that lacks the polymorphic restriction site R2, and so contains a single restriction fragment that hybridizes with the probe. The DNA in lane 2 contains the polymorphic site and so yields two fragments that hybridize with the probe. Reproduced from Brown with permission of BIOS Scientific Publishers Ltd, Oxford.

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References

Amasino RM (1986) Acceleration of nucleic acid hybridization rate by polyethylene glycol. Analytical Biochemistry 152: 304–307.

Bittner M, Kupferer P and Morris CF (1980) Electrophoretic transfer of proteins and nucleic acids from slab gels to diazobenzyloxymethyl cellulose or nitrocellulose sheets. Analytical Biochemistry 102: 459–471.

Brown TA (1999) Genomes. Oxford: BIOS Scientific Publishers.

Donis‐Keller H, Green P, Helms C et al. (1987) A genetic map of the human genome. Cell 51: 319–337.

Southern EM (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. Journal of Molecular Biology 98: 503–517.

Further Reading

Ausubel FM, Brent R, Kingston RE et al. (eds) (1993) Current Protocols in Molecular Biology. New York: John Wiley.

Brown TA (1995) Gene Cloning: An Introduction, 3rd edn. London: Chapman & Hall.

Brown TA (1998) Molecular Biology Labfax. London: Academic Press.

Dyson NJ (1991) Immobilization of nucleic acids and hybridization analysis. In: Brown TA (ed.) Essential Molecular Biology: A Practical Approach, vol. 2, pp. 111–156. Oxford: Oxford University Press.

Sambrook J, Fritsch EF and Maniatis T (1989) Molecular Cloning. A Laboratory Manual. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.

Watson JD, Gilman M, Witkowski J and Zoller M (1992) Recombinant DNA, 2nd edn. New York: WH Freeman.

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How to Cite close
Brown, Terence A(Apr 2001) Southern Blotting and Related DNA Detection Techniques. In: eLS. John Wiley & Sons Ltd, Chichester. http://www.els.net [doi: 10.1038/npg.els.0000996]