FAQ: Do you have any suggestions on how to improve DNA assembly when using synthesized DNA (e.g. gBlocks) and NEBuilder HiFi DNA Assembly Master Mix (E2621/E5520/E2623)?

Introducing an amplification step in the workflow can yield better results. Amplification of the gBlocks using a high fidelity polymerase such as Q5 (M0491S) as a first step in the assembly workflow can increase the amount of DNA available for assembly, and consequently the number of correctly assembled plasmids.