Processing
Frequently asked questions
Our Scientific Applications Support team has assembled a list of frequently asked questions to help you find answers quickly. Filter using one or more categories to focus on specific topics, or use the search bar to perform a text search.
Is the PAM sequence part of the sgRNA sequence construct?
The PAM sequence is located on the non-complementary strand. In other words, it is on the strand of DNA that contains the same DNA sequence as the target sgRNA [1]. The PAM sequence should not be included in the design of the sgRNA.
.png?sfvrsn=58c63b07_0 "pam-site-callout (1)")
.png?sfvrsn=58c63b07_0 "pam-site-callout (1)")
The most commonly used Cas9 nuclease, derived from S. pyogenes, recognizes a PAM sequence of NGG that is found directly downstream of the target sequence in the genomic DNA, on the non-target strand.
References:
- Anders C, Niewoehner O, et al. (2014) Structural basis of PAM-dependent target DNA recognition by the Cas9 endonuclease. Nature. 513(7519):569–573.