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 Citations (Scopus)

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 - Jan 1 1991
Externally publishedYes

Fingerprint

DNA Restriction Enzymes
Electrophoresis
Gels
DNA
Deconvolution
Charge coupled devices
Imaging systems
Digestion
Detectors
Equipment and Supplies
Molecules
Composite materials

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Quantitative gel electrophoresis of DNA : Resolution of overlapping bands of restriction endonuclease digests. / Ribeiro, Eldred A.; Sutherland, John C.

In: Analytical Biochemistry, Vol. 194, No. 1, 01.01.1991, p. 174-184.

Research output: Contribution to journalArticle

@article{c8f8f5d66a184f91805edaa990700247,
title = "Quantitative gel electrophoresis of DNA: Resolution of overlapping bands of restriction endonuclease digests",
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.",
author = "Ribeiro, {Eldred A.} and Sutherland, {John C.}",
year = "1991",
month = "1",
day = "1",
doi = "10.1016/0003-2697(91)90165-P",
language = "English (US)",
volume = "194",
pages = "174--184",
journal = "Analytical Biochemistry",
issn = "0003-2697",
publisher = "Academic Press Inc.",
number = "1",

}

TY - JOUR

T1 - Quantitative gel electrophoresis of DNA

T2 - Resolution of overlapping bands of restriction endonuclease digests

AU - Ribeiro, Eldred A.

AU - Sutherland, John C.

PY - 1991/1/1

Y1 - 1991/1/1

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=0025868991&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0025868991&partnerID=8YFLogxK

U2 - 10.1016/0003-2697(91)90165-P

DO - 10.1016/0003-2697(91)90165-P

M3 - Article

C2 - 1651063

AN - SCOPUS:0025868991

VL - 194

SP - 174

EP - 184

JO - Analytical Biochemistry

JF - Analytical Biochemistry

SN - 0003-2697

IS - 1

ER -