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  • Title: Quantitative gel electrophoresis of DNA: resolution of overlapping bands of restriction endonuclease digests.
    Author: Ribeiro EA, Sutherland JC.
    Journal: Anal Biochem; 1991 Apr; 194(1):174-84. PubMed ID: 1651063.
    Abstract:
    Accurate quantitation of the distribution of DNA digested to completion by a restriction endonuclease and separated in an electrophoretic gel is greatly facilitated by electronic imaging systems employing charge-coupled device detectors. Such quantitation permits the extraction of more accurate values of the lengths of both the parent molecular species and the fragments produced by a restriction endonuclease. We describe a method that determines the number of components in bands containing two or more daughter fragments that are of similar length and hence not resolved by gel electrophoresis. We also report a constrained deconvolution procedure based on the equal number of daughter molecules in a terminal digestion by a restriction endonuclease that resolves the distribution of each of the overlapping species in a composite band on a gel and hence increases the accuracy of the lengths assigned to each.
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