Requirements for a Successful ICE Analysis

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)