Top Questions

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 3NLS: 40–70 bp
    • Alt-R Cas9 H840A Nickase 3NLS: 50–70 bp
  • PAM-out orientation (right image)
    (light blue = genomic DNA, dark blue = protospacer, and orange = PAM sequence):
  • Pam site orientation (Cas9 nickase experiments)

For additional information, refer to the application note at (Support section).
  • Alt-R
  • Cas9
Product Support Topics