Oligonucleotide Annealing and sgRNA/CRISPR Vector Digestion
1:51
Identification of the Correct Recombinant Plasmids by PCR
2:42
Dual-luciferase Detection
4:16
Results: Design of sgRNA to Target Sheep DKK2 Exon 1
5:20
Conclusion
Transcrição
This protocol describes the key steps in the generation and optimization of efficient sgRNA vectors. Our preacceleration of candidate sgRNA targets relative time and inference. The main advantage of this protocol is that it makes it possible to an
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Here, we present a protocol describing a streamlined method for the efficient generation of plasmids expressing both the CRISPR enzyme and associated single guide RNA (sgRNAs). Co-transfection of mammalian cells with this sgRNA/CRISPR vector and a dual luciferase reporter vector that examines double-strand break repair allows evaluation of knockout efficiency.