Quantitative gel electrophoresis of DNA: Resolution of overlapping bands of restriction endonuclease digests

Eldred A. Ribeiro, John C. Sutherland

Research output: Contribution to journalArticle

7 Scopus citations

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 lenghts 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.

Original languageEnglish (US)
Pages (from-to)174-184
Number of pages11
JournalAnalytical Biochemistry
Volume194
Issue number1
DOIs
StatePublished - Apr 1991
Externally publishedYes

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Fingerprint Dive into the research topics of 'Quantitative gel electrophoresis of DNA: Resolution of overlapping bands of restriction endonuclease digests'. Together they form a unique fingerprint.

  • Cite this