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Protocol

An Improved Method for the Isolation of Extrachromosomal DNA from the Pathogenic Free-Living Amoeba Naegleria fowleri

by
Colm P. Roster
and
James C. Morris
*
Eukaryotic Pathogens Innovation Center, Department of Genetics and Biochemistry, Clemson University, Clemson, SC 29634, USA
*
Author to whom correspondence should be addressed.
Methods Protoc. 2026, 9(4), 105; https://doi.org/10.3390/mps9040105
Submission received: 18 May 2026 / Revised: 30 June 2026 / Accepted: 2 July 2026 / Published: 7 July 2026
(This article belongs to the Section Molecular and Cellular Biology)
Review Reports Versions Notes

Abstract

The pathogenic free-living amoeba Naegleria fowleri is the cause of primary amebic meningoencephalitis (PAM), a central nervous system infection that is almost always lethal. One of the unusual features of the amoebae is the presence of ~4000 copies of a nucleolar-localized closed circular extrachromosomal ribosomal DNA element (CERE) that encodes the cell’s ribosomal RNA repertoire. It has historically been challenging to purify large quantities of CERE, limiting our understanding of the nucleic acid. Here, we describe a methodology for CERE purification that improves yield, reduces processing times, and maintains the integrity of the plasmid. This approach will enable the study of this unique DNA architecture, advancing our understanding of the pathobiology of the organism.
Keywords: circular extrachromosomal ribosomal DNA element; episome; free-living pathogenic amoebae; Naegleria fowleri circular extrachromosomal ribosomal DNA element; episome; free-living pathogenic amoebae; Naegleria fowleri

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MDPI and ACS Style

Roster, C.P.; Morris, J.C. An Improved Method for the Isolation of Extrachromosomal DNA from the Pathogenic Free-Living Amoeba Naegleria fowleri. Methods Protoc. 2026, 9, 105. https://doi.org/10.3390/mps9040105

AMA Style

Roster CP, Morris JC. An Improved Method for the Isolation of Extrachromosomal DNA from the Pathogenic Free-Living Amoeba Naegleria fowleri. Methods and Protocols. 2026; 9(4):105. https://doi.org/10.3390/mps9040105

Chicago/Turabian Style

Roster, Colm P., and James C. Morris. 2026. "An Improved Method for the Isolation of Extrachromosomal DNA from the Pathogenic Free-Living Amoeba Naegleria fowleri" Methods and Protocols 9, no. 4: 105. https://doi.org/10.3390/mps9040105

APA Style

Roster, C. P., & Morris, J. C. (2026). An Improved Method for the Isolation of Extrachromosomal DNA from the Pathogenic Free-Living Amoeba Naegleria fowleri. Methods and Protocols, 9(4), 105. https://doi.org/10.3390/mps9040105

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