What insert sources are compatible with In-Fusion?

The following types of inserts are compatible with In?Fusion Cloning: -PCR-amplified DNA fragments carrying overhangs complementary with the termini of the adjacent DNA fragment(s), synthetic oligos, or linear vector. -DNA fragments generated by restriction digest with one or more restriction enzymes. In this instance, the required 15-bp homology must be carried by the adjacent DNA fragment(s), [...]

What are vector linearization and purification options for In-Fusion?

Linearization options include: 1. Restriction digest with one or more restriction enzymes. -For efficient In?Fusion Cloning, integrity of the linearized vector termini is essential. We recommend using high-quality restriction enzymes and performing digests over several hours. However, overnight restriction digest is not advisable. -Dephosphorylation of the vector termini is not required; the [...]

What's the allowed vector size for In-Fusion?

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, cosmids at 46 kb*) in a single reaction, without intermediate cloning into transfer/shuttle vectors. (Please see Figures 1, 2, 5, and Table III of the Adeno-X Adenoviral System 3 Brochure for details.) *The 46-kb cosmid vector, assembled by In-Fusion [...]

What are the compatible vectors for In-Fusion?

Any linear vector can be used for In-Fusion Cloning, regardless of size, composition, or available restriction site(s). The cloning reaction is followed by the rescue of a circular recombinant construct in E. coli, but please note that circular vectors are not compatible with the cloning reaction itself. Additionally, In-Fusion Cloning does not allow the assembly of covalently linked linear DNA [...]

How should I plan my In-Fusion experiment

In-Fusion Snap Assembly master mixes are available in a lyophilized (EcoDry) or liquid format. Each format also contains reagents for control experiments (linearized vector, 2-kb insert). There are two decisions to make when deciding on a kit format. First, do you want a lyophilized or liquid kit format and second, are you interested in other products to increase your cloning efficiency [...]