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RNP preparation takes 3–4 days (including Cas9 protein purification). RNP validation in vitro and in protoplasts takes 3–4 days (not including the time for plant preparation). RNP coating and delivery can be completed within 4 h. After this, the plantlets regenerate in 6–8 weeks. Identification of mutants by PCR-RE and DNA sequencing takes 3 days.
