In-Fusion Cloning tutorials

Our In‑Fusion Cloning Primer Design Tool lets you quickly and effortlessly plan out any seamless cloning project using In‑Fusion Cloning technology. The online tool is as flexible as In‑Fusion Cloning itself, accommodating:

  • single- or multiple-insert cloning without scar sequences,
  • vector linearization by inverse PCR or restriction digest, and
  • site-directed mutagenesis (insertions, deletions, substitutions).

Each step is marked for easy addition of your sequences and project specifications. Map and sequence panes let you visualize your vector as you go, and results include a final map and output protocol with oligo and PCR information. The following tutorials guide you through each step of various In‑Fusion Cloning applications. General information and help are also available below the tutorials.

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Step-by-step tutorials

Cloning one or more fragments into your final vector

Design primers to seamlessly clone single or multiple inserts directly into your destination vector at any locus.

Clone fragment(s)

Deleting a specific sequence from any vector

Design primers to delete nucleotides from your vector of choice at any locus.

Delete sequence

Inserting a specific sequence in any vector

Design primers to add nucleotides, like small tags or domains, to your vector of choice at any locus.

Insert sequence

Deleting and replacing a sequence in any vector

Design primers to simultaneously delete and replace nucleotides (e.g., for domain swapping studies) in your vector of choice at any locus.

Replace sequence


This video will show you how to design In-Fusion-ready double-stranded DNA fragments to seamlessly clone into your vector. We use the pEmpty Vector and the vectors included within the Lenti-X SARS-CoV-2 Spike Vector Set, which are included in the Lenti-X SARS-CoV-2 Packaging Single Shots (Universal) kit, as an example.