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Briefly, sgRNA complexes were synthesized by mixing equal volumes of crRNA (2 µl, 160 µM in nuclease-free water, aatcttttctgagcgcacct) and tracrRNA (2 µl, 160 µM) and incubated at 37 °C for 30 min before Cas9 (2.7 µl, 10 µg µl −1

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But before we discuss the specifics of how Perfect Amino works, we first need to cover some essential details, including information on amino acids in general