Molecular detrapping and band narrowing with high frequency modulation of pulsed field electrophoresis

In high electric fields, megabase DNA fragments are found to be trapped, i.e. to enter or migrate in the gel only very slowly, if at all, leading to very broad electrophoretic bands and loss of separation. As a consequence, low electric fields are usually used to separate these molecules by pulsed f...

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Bibliographic Details
Published in:Nucleic acids research 1990-02, Vol.18 (3), p.569-575
Main Authors: TURMEL, C, BRASSARD, E, SLATER, G. W, NOOLANDI, J
Format: Article
Language:English
Subjects:
Biological and medical sciences
Candida - genetics
Chromatin. Chromosome
DNA, Fungal - isolation & purification
Electricity
Electrophoresis - methods
Fundamental and applied biological sciences. Psychology
Molecular and cellular biology
Molecular genetics
Saccharomyces cerevisiae - genetics
Schizosaccharomyces - genetics
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Summary:In high electric fields, megabase DNA fragments are found to be trapped, i.e. to enter or migrate in the gel only very slowly, if at all, leading to very broad electrophoretic bands and loss of separation. As a consequence, low electric fields are usually used to separate these molecules by pulsed field electrophoretic methods. We report here that high-frequency pulses eliminate the molecular trapping found in continuous fields. When high frequency pulses are used to modulate the longer pulses used in pulsed field electrophoresis, narrower bands result, and higher fields can be used. We suggest that this is due to effects that occur on the length scale of a single pore.
ISSN:0305-1048
1362-4962
DOI:10.1093/nar/18.3.569