Protocol

Alkaline Agarose Gel Electrophoresis

Abstract

Alkaline agarose gels are run at high pH, which causes each thymine and guanine residue to lose a proton and thus prevents the formation of hydrogen bonds with their adenine and cytosine partners. The denatured DNA is maintained in a single-stranded state and migrates through an alkaline agarose gel as a function of its size. Other denaturants such as formamide and urea do not work well because they cause the agarose to become rubbery.

Footnotes

  • From the Molecular Cloning collection, edited by Michael R. Green and Joseph Sambrook.

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