Make Gibson Assembly 2X mastermix v1

Gibson assembly, a homology-based cloning method, enables much simpler and more efficient cloning compared to traditional T4 ligation. The commercially available product based on this method is the NEBuilder‱ HiFi DNA Assembly Master Mix (NEB, E2621), which has also been widely used in many labs. This protocol follows the method refined by Rabe et al. (2020), and is directly based on the procedure summarized by Julian Wills from the David Liu lab. The key point of this protocol is the addition of SSB to protect ssDNA from endonuclease activity. The homemade Gibson mix is about 6.5 times cheaper than the NEBuilder HiFi mix.