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Purification and functional validation of LtCas12a protein

Bo Zhou,Ye Chen,Lifang Li,Jiashuo Liu,Yuyan Wang,Zheying Huang,Zheng Hu,Rui Tian,Zhen Li

Summary Here, we present a protocol for generating LtCas12a protein recognizing distinct TTNA (N represented A, T, C, G) protospacer adjacent motif sequence. We describe steps for transforming and harvesting bacterial cells and protein purification including nickel affinity chromatography and dialysis. We then detail procedures for verification of LtCas12a with cis- and trans-cleavage activities. For complete details on the use and execution of this protocol, please refer to Chen et al. (2023). 1