For point mutation, it is suggested to use asymmetric ssDNA design. You can read this paper for more design tips. For large gene insertion, homology arms with 300-1000 bp flanking the insertion gene have been reported. In most cases, 500 bp long homology arms should work. It is important to design the ssDNA template containing a silent mutation to mutate the sgRNA PAM sequence in order to avoid secondary cleavage. KI donor design can be complicated. It is highly suggested to use a software (e.g. snapgene) to view and edit the sequence.
Articles in this section
- What is the process of developing GenWandTM dsDNA?
- Why should I choose GenWandTM dsDNA over PCR dsDNA?
- Do I need to worry about dsDNA contamination in the GenExactTM ssDNA product?
- How do you purify the final GenExactTM ssDNA product?
- Why do you do double-sequencing, including the final GenExactTM ssDNA product?
- What are the quality control tests you provide for the GenExactTM ssDNA templates?
- What is the maximum GenExactTM ssDNA length GenScript produces?
- What is the maximum quantity GenScript provides for GenWandTM dsDNA and GenExactTM ssDNA?
- How should I choose between GenExactTM ssDNA and GenWandTM dsDNA?
- What are the applications of HDR templates (GenExactTM ssDNA and GenWandTM dsDNA)?