&lt;title&gt;Dispersion functions and factors that determine resolution for DNA sequencing by gel electrophoresis&lt;/title&gt;

Sutherland, John C.; Reynolds, Kiley J.; Fisk, David J.

doi:10.1117/12.237621

Cited by 2 publications

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<title>Linking electrophoretic resolution with experimental conditions</title>

Sutherland

1997

SPIE Proceedings

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Previously, we described a quantitative measure of electrophoretic resolution called Resolving Power that can be computed for individual bands of DNA (or other molecules) separated by electrophoresis. An alternate approach is to determine analytical functions, based on a few experimentally determined parameters, that describe the resolving power of a particular electrophoretic system for all of the length classes of molecules that are separated. Such analytical functions have been obtained for singlestranded DNA separated in a polyacrylamide gel and detected at a fixed distance from the origin of electrophoresis. Six experimentally determined constants are required to describe the analytical function for resolving power in this system: four constants describe the mobility of DNA in the gel as a function of molecular length (or time of arrival at the detector), and two describe the width of the bands as a function of the length of molecular length (or time of arrival at the detector).One of our goals is to develop systematic methods of improving the resolving power of longer molecules, and hence extending the number of bases that can be determined in a single sequencing experiment. Our approach is to determine how the small number ( 6) of parameters that describe resolving power depend on experimentally controllably conditions (e. g. gel composition, electric field strength, temperature ...). Knowing such relationships should permit systematic selection of combinations of experimental conditions that improve resolving power.Here, we show the relationship between the four parameters describing DNA mobility as a function of molecular length (mass) for systematic variations of gel composition and electric field strength. The data set employed is for double-stranded DNA separated in agarose gels, but the principles are similar to those encountered in DNA sequencing studies.

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<title>Linking electrophoretic resolution with experimental conditions</title>

Sutherland

1997

SPIE Proceedings

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DNA Damage Quantitation by Alkaline Gel Electrophoresis

Sutherland

Bennett

Sutherland

Methods in Molecular Biology

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Quantifying DNA lesions provides a powerful way to assess the level of endogenous damage or the damage level induced by radiation, chemical or other agents, as well as the ability of cells to repair such damages. Quantitative gel electrophoresis of experimental DNAs along with DNA length standards, imaging the resulting dispersed DNA and calculating the population average length allows accurate measurement of lesion frequencies. Number average length analysis provides high sensitivity and does not require any specific distribution of lesions within the DNA molecules. These methods are readily applicable to strand breaks and ultraviolet radiation induced pyrimidine dimers, but can also be used-with appropriate modifications-for ionizing radiation-induced lesions such as oxidized bases and abasic sites.

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<title>Dispersion functions and factors that determine resolution for DNA sequencing by gel electrophoresis</title>

Cited by 2 publications

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<title>Linking electrophoretic resolution with experimental conditions</title>

<title>Linking electrophoretic resolution with experimental conditions</title>

DNA Damage Quantitation by Alkaline Gel Electrophoresis

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