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.
CITATION STYLE
Green, M. R., & Sambrook, J. (2021). Alkaline agarose gel electrophoresis. Cold Spring Harbor Protocols, 2021(11), 458–462. https://doi.org/10.1101/pdb.prot100438
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