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DOI: 10.1186/s13007-017-0170-x

A robust protocol for efficient generation, and genomic characterization of insertional mutants of Chlamydomonas reinhardtii

Steve V. Pollock, Bratati Mukherjee, Joanna Bajsa-Hirschel, Marylou C. Machingura, Ananya Mukherjee, Arthur R. Grossman, and James V. Moroney

Abstract

Random insertional mutagenesis of Chlamydomonas reinhardtii using drug resistance cassettes has contributed to the generation of tens of thousands of transformants in dozens of labs around the world. In many instances these insertional mutants have helped elucidate the genetic basis of various physiological processes in this model organism. Unfortunately, the insertion sites of many interesting mutants are never defined due to experimental difficulties in establishing the location of the inserted cassette in the Chlamydomonas genome. It is fairly common that several months, or even years of work are conducted with no result. Here we describe a robust method to identify the location of the inserted DNA cassette in the Chlamydomonas genome.

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