Zhang Lab's CRISPR Frequently Asked Questions
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Collection
...There is no need to add the NGG PAM sequence. Because the 'NGG' of the PAM is used to select your genomic...target site sequence (protospacer+PAM) is located? If there are other PAMs in the region, will they also ... the PAM (my target sequence) in my HR template an issue? Yes. If you have intact protospacer+PAM sequence...good way is to mutate the PAM 'NGG' sequence in the HR template. Since the PAM sequence is required for... this Nature Protocol article . Should I add the PAM sequence to the oligo when cloning my target spacers...be targeted? The Cas9 cuts 3-4bp upstream of the PAM sequence. There can be some off-target DSBs using... (gRNA). These considerations only matter if the PAM sequence is immediately adjacent to the nearly-homologous...