...determination (CFD score) is used to measure specificity of a guide to the target. The CFD score ranges ...performed because weak gRNAs (CFD < 20) bind almost not at all, and strong gRNAs (CFD > 80) bind almost as much...much as perfect gRNAs (CFD = 100). CFD scores (which range between 0 and 100) are then converted to a ... best practices. For example, it ranks guides by CFD score, and it mutates HDR donor templates to avoid... formula that does just this: Rank = (compressed CFD score) * (gaussian normalized cut-to-insert distance...mismatches between the guide and the DNA target. CFD scores are then modified (“transformed”) such that... PAM) in the target region after HDR. If the max CFD score is less than 0.03, it stops mutating. (This...