Denaturing Gel Electrophoresis of DNA in Alkaline Agarose Gels

Abstract

One of the simplest ways to separate DNA under denaturing conditions is to use an alkaline agarose gel. This method is not, however, suitable for RNA because it is rapidly hydrolysed in alkaline conditions. Similarly, DNA containing ribonucleotides will be nicked.

Keywords: RNA/DNA; denature; alkaline agarose

Figure 1.

Submarine slab gel apparatus.

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References

Bailey JM and Davidson N (1976) Methylmercury as a reversible denaturing agent for agarose gel electrophoresis. Analytical Biochemistry 70: 75.

Fangman WL (1978) Separation of very large DNA molecules by gel electrophoresis. Nucleic Acids Research 5: 653.

Jovin TM (1971) Methods in Enzymology 21: 179.

McDonell MW, Simon MN and Studier FW (1977) Analysis of restriction fragments of T7 DNA and determination of molecular weights by electrophoresis in neutral and alkaline gels. Journal of Molecular Biology 110: 119.

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Southern, Edwin M(Oct 2002) Denaturing Gel Electrophoresis of DNA in Alkaline Agarose Gels. In: eLS. John Wiley & Sons Ltd, Chichester. http://www.els.net [doi: 10.1038/npg.els.0003758]