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The plasmids encoding PE2 (Plasmid#132775), as well as the plasmid expressing the pegRNA (Plasmid #132777), were obtained from Addgene. PegRNAs were cloned into the plasmid backbone with paired oligonucleotides (IDT, Supplementary Table S1), as previously described (Anzalone et al., 2019). Briefly, the oligonucleotides used to create the guide sequences were hybridized, phosphorylated, and cloned into the sgRNA vector using BsaI-HFv2 (NEB), in a reaction that included T4-PNK (NEB) and T4 DNA Ligase (NEB).
Copyright and License information: ©2023 Kim, Yuan, Woods, Newton, Perez-Pinera and Song.
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