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 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.
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:

  1. Anders C, Niewoehner O, et al. (2014) Structural basis of PAM-dependent target DNA recognition by the Cas9 endonuclease. Nature. 513(7519):569–573.

Categories:
CRISPR genome editing

Tags:
crispr alt-r cas9