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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.
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Is the PAM sequence part of the gRNA sequence construct?
The PAM sequence is located on the non-complementary strand of the target DNA. In other words, it is on the strand of DNA that contains the same DNA sequence as the sgRNA [1]. The PAM sequence should not be included in the design of the sgRNA.
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.png?sfvrsn=58c63b07_0 "pam-site-callout (1)")
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.
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References:
1. Anders C, Niewoehner O, Duerst A, Jinek M. Structural basis of PAM-dependent target DNA recognition by the Cas9 endonuclease. Nature. 2014;513(7519):569-573.