Abstract: The present disclosure generally relates to compositions and methods for the synthesis of nucleic acid molecules with low error rates. Provided, as examples, are compositions and methods for high throughput synthesis and assembly of nucleic acid molecules, in many instances, with high sequence fidelity. In many instances, thermostable mismatch recognition proteins (e.g., thermostable mismatch binding protein, thermostable mismatch endonucleases) will be present in compositions and use methods provided.

Abstract: Compositions and methods are provided herein for high throughput synthesis and assembly of nucleic acid molecules. Specific aspects include methods and rules for designing, grouping and pooling of nucleic acid molecules for efficient multiplex assembly, amplification, processing and analysis to obtain error-free assembly products. Provided compositions and methods allow for miniaturization, parallelization, high throughput production and scaling, and cost reduction in gene synthesis workflows.