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:
- Alt-R Cas9 D10A Nickase V3: 40–70 bp
- Alt-R Cas9 H840A Nickase V3: 50–70 bp
- PAM-out orientation (right image)
(light blue = genomic DNA, dark blue = protospacer, and orange = PAM sequence):
For HDR designs, we recommend using our HDR design tool which incorporates these design recommendations.