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Fig. 1 | Plant Methods

Fig. 1

From: Combined fluorescent seed selection and multiplex CRISPR/Cas9 assembly for fast generation of multiple Arabidopsis mutants

Fig. 1

CRISPR/Cas cloning strategy. a Oligo annealing-based cloning of chosen gRNAs into pRU41-48 entry vectors containing pU6 and pU3 promoters. b, c Golden Gate assembly of up to eight gRNAs into corresponding intermediate vectors containing attL1-attL2 for single Gateway LR reaction. d Final single Gateway assembly into T-DNA vectors containing PcUBi4-2 or pEC1.2 promoters driving Cas9 of interest. The vectors contain one of three fluorescent seed selection cassettes, FastRed, FastGreen and FastCyan

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Plant Methods

ISSN: 1746-4811