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Malar J. 2012 Jun 21;11:210. doi: 10.1186/1475-2875-11-210.

Comparison of the absolute and relative efficiencies of electroporation-based transfection protocols for Plasmodium falciparum.

Author information

1
Institute for Science and Technology in Medicine, Keele University, Staffordshire, ST5 5BG, UK.

Abstract

BACKGROUND:

Several electroporation protocols exist to transfect exogenous DNA into Plasmodium falciparum. To date, however, only a subjective analysis of their relative efficiencies has been reported.

METHODS:

A time-course of luciferase reporter expression is used to provide an objective quantitative analysis of the absolute efficiency of three electroporation techniques; direct electroporation of ring stage infected erythrocytes, preloading of erythrocytes and a novel "double-tap" protocol that combines both approaches.

RESULTS:

Preloading of erythrocytes shows a mean efficiency of 9.59 x 10(-6), some 5-180 fold more efficient than matched experiments utilizing the "double-tap" and direct electroporation of ring stage infected erythrocytes alone, respectively.

CONCLUSION:

Evidence presented here provides the first quantitative assessment of both the absolute and relative efficiencies of a key molecular tool used to study the biology and pathogenesis of this important human pathogen.

PMID:
22720754
PMCID:
PMC3407700
DOI:
10.1186/1475-2875-11-210
[Indexed for MEDLINE]
Free PMC Article

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