We recommend incubating the genomic DNA and Cas9 RNPs at 37 ºC for 15 minutes.

Shorter times could result in incomplete cleavage. While longer times up to 60 minutes may be used, this could result in an increased number of off-target but not on-target reads.

A serial testing should be performed on the sample of interest should users choose to use longer or short cleavage time than the recommended 15 minutes.