Enabling one-pot Golden Gate assemblies of unprecedented complexity using data-optimized assembly design.

ABSTRACT: DNA assembly is an integral part of modern synthetic biology, as intricate genetic engineering projects require robust molecular cloning workflows. Golden Gate assembly is a frequently employed DNA assembly methodology that utilizes a Type IIS restriction enzyme and a DNA ligase to generate recombinant DNA constructs from smaller DNA fragments. However, the utility of this methodology has been limited by a lack of resources to guide experimental design. For example, selection of the DNA sequences at fusion sites between fragments is based on broad assembly guidelines or pre-vetted sets of junctions, rather than being customized for a particular application or cloning project. To facilitate the design of robust assembly reactions, we developed a high-throughput DNA sequencing assay to exami

SUBMITTER: Pryor JM 

PROVIDER: S-EPMC7467295 | biostudies-literature | 2020

REPOSITORIES: biostudies-literature