Fig. 2.

Fig. 2. Cleavage of linearized or circular dsDNA by CRISPR/Cas9 in vitro. (A) Designed sgRNA and linearized pX330 by EcoRI (200 ng/μl, 8.5 kb) cleaved by Cas9 endonuclease. Lane M: marker; Lanes 1 and 2: linearized pX330 cleaved by Cas9 at 37°C for 2 h and overnight, respectively. (B) Designed sgRNAs and circular pX330 (200 ng/μl, 8.5 kb) cleaved by Cas9 endonuclease. Lane M: marker; Lanes 1-4: circular pX330 cleaved by Cas9 and sgRNA at different concentrations of 60 nM, 30 nM, 15 nM and 120 nM. (C) Designed sgRNAs and circular pYPZ37 (415 ng/μl, 28.0 kb) cleaved by Cas9 endonuclease. Lane M: marker; Lane 1-5: circular pYPZ37 cleaved by Cas9 at different amounts of 1,000 ng, 500 ng, 200 ng, 100 ng, and 50 ng.

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