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Fig. 2 | Plant Methods

Fig. 2

From: A simple and efficient CRISPR/Cas9 platform for induction of single and multiple, heritable mutations in barley (Hordeum vulgare L.)

Fig. 2

PCR/RE screening of T0 plants transformed with Cas9:sgRNA constructs for simplex editing of HvCKX1 (a), Nud (b), and the PTG construct for multiplex editing of the HvCKX1 and HvCKX3 genes (c). PCR products of the appropriate target gene were treated by restriction enzymes that overlap the potential mutation site (indicated above the aligned sequences). Red arrowheads indicate uncut bands of amplicons with mutations; black arrowheads indicate bands of wild-type sequences cut by an enzyme. PCR products from selected samples were cloned and sequenced to identify the pattern of mutations; target sequences for sgRNA:Cas9 are marked in yellow, PAM motifs are marked in light blue; deletions are indicated by dashes and insertions by red letters

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