Pulsed-field gel electrophoresis of circular DNA

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Pulsed-field gel electrophoresis of circular DNA.

Mobility of supercoiled (form I) and nicked circular (form II) plasmid DNAs was determined on two major forms of pulsed-field electrophoresis, CHEF and OFAGE. Plasmids with molecular lengths ranging from 2.30 to 17.8 kilobase pairs (kb) were used with Saccharomyces cerevisiae chromosomes as standards. Agarose gel concentrations were varied from 0.3 to 2.0 percent, with higher percentage gels re...

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Pulsed-field gel electrophoresis.

DNA molecules in an agarose matrix elongate and migrate toward the anode when exposed to an electric field. An agarose matrix represents a highly irregular network with pores of various dimensions, large open areas, and regions with different densities. During electrophoresis, migration will be retarded by obstructions that prevent sieving by size and result in "end-on" migration as if through ...

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Estimation of circular DNA size using gamma-irradiation and pulsed-field gel electrophoresis.

A method is described for estimating the size of large circular DNAs found within complex chromosomal DNA preparations. DNAs are treated with low levels of gamma-irradiation, sufficient to introduce a single double-stranded break per circle, and the resulting linear DNA is sized by pulsed-field electrophoresis and blot hybridization. The method is fast, reproducible, and very conveniently appli...

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Separations of open-circular DNA using pulsed-field electrophoresis.

The effect of high electric fields on the gel-electrophoretic mobility of open-circular DNA in agarose differs dramatically from that on linear molecules of the same molecular weight. At high fields, sufficiently large circular forms are prevented from migrating into the gel whereas linear molecules and smaller circular DNAs migrate normally. This effect is strongly field dependent, affecting c...

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Pulsed-field gel electrophoresis of very large DNA molecules.

FACTORS THAT AFFECT PFG SEPARATIONS........................................................................ 294 Molecular Properties ............................................... .................................... ............. 294 Pulse Times............................................................................................................... 295 Electrical Field Shape ..............

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ژورنال

عنوان ژورنال: Nucleic Acids Research

سال: 1989

ISSN: 0305-1048,1362-4962

DOI: 10.1093/nar/17.11.4359