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Published in: Malaria Journal 1/2012

Open Access 01-12-2012 | Methodology

Plate-based transfection and culturing technique for genetic manipulation of Plasmodium falciparum

Authors: Florence Caro, Mathew G Miller, Joseph L DeRisi

Published in: Malaria Journal | Issue 1/2012

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Abstract

Genetic manipulation of malaria parasites remains an inefficient, time-consuming and resource-intensive process. Presented here is a set of methods for 96-well plate-based transfection and culture that improve the efficiency of genetic manipulation of Plasmodium falciparum. Compared to standard protocols plate-based transfection requires 20-fold less DNA, transient transfection efficiency achieved is approximately seven-fold higher, whilst stable transfection success rate is above 90%. Furthermore the utility of this set of protocols to generate a knockout of the PfRH3 pseudogene, screened by whole-cell PCR, is demonstrated. The methods and tools presented here will facilitate genome-scale genetic manipulation of P. falciparum.
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Metadata
Title
Plate-based transfection and culturing technique for genetic manipulation of Plasmodium falciparum
Authors
Florence Caro
Mathew G Miller
Joseph L DeRisi
Publication date
01-12-2012
Publisher
BioMed Central
Published in
Malaria Journal / Issue 1/2012
Electronic ISSN: 1475-2875
DOI
https://doi.org/10.1186/1475-2875-11-22

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