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Identifying and isolating meiotic mutants in a polyploid brassica crop

Abstract : This chapter provides a detailed description of TILLING and CRISPR-Cas9 approaches for the purpose of studying genes/factors involved in meiotic recombination in the polyploid species B. napus. The TILLING approach involves the screening and identification of EMS-mutagenized M2 B. napus plants. The strategy for high-throughput plant pooling, the set up for microfluidic PCR and sequencing is provided and the parameters for the analysis of sequence results and the detection of mutants are explained. The CRISPR-Cas system relies on the optimal design of guide RNAs and their efficient expression. The procedure for the generation and detection of knockout mutants is described with the aims to simultaneously target homologous genes.
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Contributor : Christine Molé Connect in order to contact the contributor
Submitted on : Wednesday, August 5, 2020 - 5:40:01 PM
Last modification on : Friday, August 5, 2022 - 2:38:11 PM



Marina Pfalz, Adrián Gonzalo, Nicolas Christophorou, Aurélien Blary, Aurélie Berard, et al.. Identifying and isolating meiotic mutants in a polyploid brassica crop. Mónica Pradillo; Stefan Heckmann. Plant Meiosis : methods and protocols, 2061, Humana, 405 p., 2020, Methods in Molecular Biology, 978-1-4939-9817-3 ; 978-1-4939-9818-0. ⟨10.1007/978-1-4939-9818-0_21⟩. ⟨hal-02912375⟩



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