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. Author manuscript; available in PMC: 2017 Jan 1.
Published in final edited form as: Curr Protoc Hum Genet. 2016 Jan 1;88:21.4.1–21.4.23. doi: 10.1002/0471142905.hg2104s88

Figure 2. gRNA Plasmid Ligation Conditions.

Figure 2

(A) Example of annealed and phosphorylated gRNA target sequence oligo with directional sticky 5′ overhangs. (B) Example of directional ligation into BbsI digested dephosphorylated pX330 vector. (C) Example of the completed sgRNA (blue/red) and how it will recognize the genomic target sequence (black) with Cas9 PAM binding motif (green).