Frequently asked questions

Our Scientific Applications Support team has assembled a list of frequently asked questions to help you find answers quickly. Filter using one or more categories to focus on specific topics, or use the search bar to perform a text search.

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How far apart should I design my two guides when using the Alt-R® nickases?

When designing crRNA pairs to use with the Alt-R nickases, we recommend the following:

  • Distance between cleavage sites:
  • PAM-out orientation (right image)
    (light blue = genomic DNA, dark blue = protospacer, and orange = PAM sequence):

cas9-pam-site-orientation-for-nickase-experiments