...and a ssDNA-fluorescently quenched reporter is added. As this reporter is degraded by Cas12a, it will release... using a fluorescent reporter. In SHERLOCK, a quenched ssRNA fluorescent reporter is added to the reaction...insertions. When designing the donor template, incorporating mutations that prevent further Cas9 cleavage...Overview of HDR. In most applications, a large portion of the Cas9-induced DSBs will still be repaired...the desired HDR-edited allele. Therefore, it is important to confirm the presence of the desired edit experimentally...pairs to the 3’ flap, and the desired edit is incorporated into the DNA by reverse transcription. The edited...applications where orientation of the insert is important. However, type I-F CASTs favor insertion in the...