Can I use a circular cloning vector for In-Fusion Cloning?

No, circular cloning vectors are not compatible with In?Fusion Cloning. A vector must be linearized via restriction digest or inverse PCR.In-Fusion Cloning [...]

Can I use In-Fusion Cloning to assemble a covalently linked linear DNA molecule?

No, In-Fusion Cloning does not allow the assembly of covalently linked linear DNA molecules. In-Fusion Cloning kit components include a linearized cloning vector, enabling the rescue of a circular recombinant construct in E. coli.In-Fusion Cloning [...]

Is In-Fusion Cloning compatible with vectors carrying repeated sequences?

Yes—our scientists routinely use In?Fusion Cloning to clone transgenes into lentiviral or retroviral vectors that carry long terminal repeats (LTRs), as well as adenoviral vectors that carry inverted terminal repeats (ITRs).In-Fusion Cloning [...]

Are large cloning vectors compatible with In-Fusion Cloning?

Yes, In-Fusion technology allows easy cloning of single or multiple DNA fragments directly into large vectors (e.g., adenoviral vectors at 32.6–36 kb) in a single reaction, without intermediate cloning into transfer/shuttle vectors.In-Fusion Cloning [...]

Do I have to dephosphorylate the termini of a linearized vector for In-Fusion Cloning?

No, dephosphorylation of the vector termini is neither required nor recommended for In-Fusion Cloning.In-Fusion Cloning [...]