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Original Articles

Fast Procedure To Distinguish Circular and Linear DNA Molecules in Pulsed Field Gel Electrophoresis

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Pages 745-753 | Received 25 Aug 1995, Accepted 14 Nov 1995, Published online: 22 Aug 2006
 

Abstract

We describe here a fast procedure to identify, by Pulsed Field Gel Electrophoresis (PFGE), circular DNA molecules, displaying shift mobility dependent on ethidium bromide (EtBr) concentration. Several identical DNA samples were coelectrophoresed in a multilane gel with different EtBr concentrations in each lane. Samples were run in a Contour Clamped Homogeneous Electric Field (CHEF) or in a small CHEF (miniCHEF) equipment for 15 or 4 h, respectively. Circular DNA molecules from the protozoan parasite Entamoeba histolytica were detected in a single running. The circles were isolated from the gels and then their structure was confirmed by transmission electron microscopy. By hybridization it was shown that the circles obtained corresponded to the ribosomal DNA episome.

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