Titlebook: CRISPR-Cas Methods; Volume 2 M. Tofazzal Islam,Kutubuddin Ali Molla Book 2021 The Editor(s) (if applicable) and The Author(s), under exclus

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書目名稱CRISPR-Cas Methods影響因子(影響力)




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Chr. Gutenbrunner,G. Hildebrandtew crop variety. This chapter briefly describes the methods of CRISPR-Cas9 vector construction with single and multiple gene targets in rice. The protocol covers all important steps of target sequence selection, single/multiple-target vector construction, mutation detection, and selection of T-DNA free mutant lines.

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In Vitro Cas9 Cleavage Assay to Check Guide RNA Efficiency,ro method to screen multiple sgRNAs to identify the most suitable one that can efficiently introduce a double-stranded break at a?particular genomic target site. This screening method allows a researcher to choose the best one among several online predicted sgRNAs prior to deliver genome editing reagents into live plant or animal cells.

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Generation of Knockout and Fragment Deletion Mutants in Soybean by CRISPR-Cas9,o the CRISPR-Cas9-mediated targeted mutagenesis or large fragment deletions in soybean. Detailed procedures will guide through the essential steps including the design of sgRNAs, construction of CRISPR-Cas9 vectors, .-mediated soybean transformation, and identification of mutant lines.

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1949-2448 ation advice from the experts.This second volume provides new and updated methods detailing advancements in CRISPR-Cas technical protocols. Chapters guide readers through protocols on prime editing, base editing, multiplex editing, editing in cell-free extract, in silico analysis of gRNA secondary s

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https://doi.org/10.1007/978-3-662-25284-0e, wheat, maize, and tomato, which substantially expands the scope and capabilities of precision plant breeding. Here, we describe a fast and efficient method for construction of prime editing vectors based on Gateway assembly and efficiency assessment of prime editors through transient expression analyses in rice protoplasts.

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