Zhang Lab's CRISPR Frequently Asked Questions
Type
Collection
...ideally less than 10bp away. For small (<50bp) changes, you should consider using ssDNA oligo as donor...80bp of homology arms on each side flanking the change you'd like to introduce. We use Ultramer oligo ...the Cas9/sgRNAs vector (like PX330). For large changes (>100bp insertions or deletions), we typically ...to mutate the PAM 'NGG' in the HR template, like change it to 'NGT', or 'NGA', or 'NGC' (if it's within...required for successful targeting and cleavage, changing it to'NGT' or 'NGC', in addition to mutations ...typically use PAGE purified long oligos. For large changes (>100bp insertions or deletions), the Zhang lab...