The Gibson Assembly ® HiFi 1 Step method utilizes a blend of enzymes to assemble the DNA quickly and efficiently. The use of a High-Fidelity DNA polymerase enzyme allowsfor error free cloning resulting in a seamless construct ready for multiple downstream applications. The HiFi 1 Step method allows Gibson Assembly of up to 5 fragments using a one tube, one-step, isothermal reaction. Multiple DNA fragments can be joined with greater than 90% cloning efficiency. Large constructs up to 100kb in length can be efficiently assembled in multi stage reactions.