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. Author manuscript; available in PMC: 2023 Aug 16.
Published in final edited form as: Methods Mol Biol. 2023;2631:155–182. doi: 10.1007/978-1-0716-2990-1_6

Figure 5: Primer design and PCR strategy for KI projects using dsDNA donor.

Figure 5:

Long PCR primers are designed to the 5ʹ and 3ʹ regions of the desired targeted integration event. Each primer set contains a Gen primer designed outside of the homology arm (blue) and a Junc primer designed inside the target KI region (red). Short PCRs primers are designed for nested PCR using the long PCR product as the template to create amplicons that are within the NGS sequence length limits. Short PCR primers consist of a Junc primer, and a HA primer designed within the homology arm. DS tags shown in yellow.