Abstract

Abstract DNA molecules longer than 25 kb are poorly resolved by standard agarose gel electrophoresis. These longer molecules can be resolved using several techniques that periodically change the direction of the electric field in the gel. This unit describes the simplest and most generally useful of the pulsed‐field techniques, field inversion electrophoresis, which can be tuned to resolve molecules from ˜10 to 2000 kb (or more with specialized equipment). To resolve molecules beyond the range of field inversion, it is necessary to use some sort of field‐angle alternation electrophoresis such as CHEF (contour‐clamped homogeneous electric field; described in an ). A method is also provided for preparing high‐molecular‐weight DNA samples and size markers embedded in agarose blocks.

Keywords

AgarosePulsed-field gel electrophoresisElectrophoresisElectric fieldAgarose gel electrophoresisGel electrophoresisGel electrophoresis of nucleic acidsChromatographyDNAMoleculeChemistryAnalytical Chemistry (journal)PhysicsBiochemistry

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Year
2000
Type
article
Volume
51
Issue
1
Pages
Unit2.5B-Unit2.5B
Citations
46
Access
Closed

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Michael Finney (2000). Pulsed‐Field Gel Electrophoresis. Current Protocols in Molecular Biology , 51 (1) , Unit2.5B-Unit2.5B. https://doi.org/10.1002/0471142727.mb0205bs51

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DOI
10.1002/0471142727.mb0205bs51