There are several requirements for successful ICE Analysis:
- Most updated version of Google Chrome to launch the ICE Analysis tool
- The correct sgRNA sequence(s) as RNA or DNA, without the PAM site
- For knock-ins: donor template with 15 bp homology arms at each end (total maximum length is 300 bp)
- High quality Sanger sequencing files (.ab1) for a wild-type (control) sample and an experimental sample that were generated with the same sequencing/PCR primers
- Sanger sequencing files for PCR products ranging from 400-800 bp with the PCR/sequencing primers being at least 150 bp away from the sgRNA cut site(s)