COMPOSITIONS AND METHODS FOR NUCLEIC ACID ASSEMBLY

摘要

The present disclosure generally relates to compositions and methods for the assembly of nucleic acid molecules into larger nucleic acid molecules. Also provided are compositions and methods for seamlessly connection of nucleic acid molecules with high sequence fidelity.

主权项

1. A method for covalently linking two nucleic acid segments, the method comprising:
(a) incubating the two nucleic acid segments with an exonuclease under conditions that allow for digestion of termini of the two nucleic acid segments to form complementary single-stranded regions on each nucleic acid segment and hybridization of the complementary single-stranded regions, wherein each of the two nucleic acid segments comprises an exonuclease resistant region within 30 nucleotides of the end of the complementary terminus, and (b) covalently connecting at least one strand of the hybridized termini formed in (a) resulting in the linkage of the two nucleic acid segments.