In-Fusion HD EcoDry Cloning Plus products provide our In-Fusion HD cloning enzyme mix in a lyophilized, ready-to-use format that can be stored at room temperature. The cloning reagent is pre-aliquoted and dried down in tubes that are covered with an aluminum seal. This convenient cloning enzyme format is stable at room temperature, making it ideal for high-throughput cloning with robotic liquid handlers. Whether at the bench or with automation platforms, use of pre-aliquoted reagents means minimal handling, eliminating chances of contamination, pipetting errors, or missing reagents. Just like the liquid format, In-Fusion HD EcoDry Cloning Plus allows you to clone any PCR fragment into any linearized vector in a single step without restriction digestion of the PCR fragment, ligation, or blunt-end polishing.

The ability to clone directly into any vector at any restriction site eliminates the need for any further subcloning or manipulation, greatly simplifying the cloning workflow and further enabling a transition to high-throughput applications. The In-Fusion HD enzyme premix fuses PCR-generated sequences and linearized vectors efficiently and precisely, utilizing a 15-bp overlap at their ends. This 15-bp overlap can be engineered by designing custom primers for amplification of the desired sequences using our primer design tool.

In-Fusion Cloning technology is based on the removal of nucleotides from the 3" end of the linear DNA strands. This allows complementary base pairs between two fragments of DNA base pairs to anneal, leading to fragment joining. This method can be used to clone single or multiple fragments into a single vector without subcloning. Additional applications include directional cDNA cloning, mutagenesis, gene synthesis, high-throughput cloning, as well as for the addition of adaptors, linkers, and protein tags.

In-Fusion HD EcoDry Cloning Plus components

• In-Fusion HD EcoDry Mix (lyophilized; store at room temperature)

  In-Fusion HD enzyme mix fuses DNA fragments efficiently and precisely by recognizing a 15-bp overlap at their ends—without the use of ligase. Provided in a dried-down format, pre-aliquoted in 8-well tube strips or 96-well plates, depending on kit size. Note: the liquid kit format includes a 5X enzyme premix in 20-μl or 100-μl aliquots, depending on kit size.

• CloneAmp HiFi PCR Premix (liquid; store at –°20C)

  CloneAmp HiFi PCR Premix is a convenient 2X liquid master mix that offers exceptionally accurate, efficient, and fast DNA amplification. The premix contains dNTPs and an optimized buffer, allows rapid set-up of PCR reactions, and facilitates successful cloning. The polymerase master mix is provided in 625-μl aliquots in both EcoDry and liquid kits.

• Stellar Competent Cells (frozen; store at –80°C)

  High-efficiency competent cells are essential to the success of In-Fusion Cloning. An E. coli HST08 strain is included that provides high transformation efficiency (greater than 5 x 10⁸ cfu/µg) and complements the efficiency of all In-Fusion HD EcoDry Cloning Plus kits. Cells are provided in 100-μl aliquots in individual tubes. The 96-rxn EcoDry kit includes the cells pre-aliquoted in 96-well plates (20 μl in each well).

• NucleoSpin Gel and PCR Clean-up kit (store at room temperature)

  For gel extraction as well as PCR purification. Provided as individual columns or 96-well plate purifications, depending on kit size.