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.
All Keywords
【저자키워드】 structural biology, Cancer, genetics, CRISPR, Protein Biochemistry, Protein expression and purification,
【저자키워드】 structural biology, Cancer, genetics, CRISPR, Protein Biochemistry, Protein expression and purification,