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Reverse genetics in Chlamydomonas: a platform for isolating insertional mutants
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Reverse genetics in Chlamydomonas: a platform for isolating insertional mutants
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Journal Article
Reverse genetics in Chlamydomonas: a platform for isolating insertional mutants
2011
Overview
A method was developed to identify insertional mutants of
Chlamydomonas reinhardtii
disrupted for selected target genes. The approach relies on the generation of thousands of transformants followed by PCR-based screenings that allow for identification of strains harboring the introduced marker gene within specific genes of interest. Our results highlight the strengths and limitations of two independent screens that differed in the nature of the marker DNA used (PCR-amplified fragment containing the plasmid-free marker versus entire linearized plasmid with the marker) and in the strategies used to maintain and store transformants.
Publisher
BioMed Central,BioMed Central Ltd,BMC
Subject
