Proofreading, error deletion, and ligation method for synthesis of high-fidelity polynucleotide sequences

Abstract

Methods and apparatuses for solid-phase oligonucleotide synthesis and forming long polynucleotides. One exemplary method includes synthesizing a sense oligonucleotide; synthesizing an antisense oligonucleotide; annealing the sense and antisense oligonucleotides to form double stranded DNA (dsDNA); capping the ends of the dsDNA; cleaving the dsDNA wherein cleavage occurs at or near a Watson-Crick base pair mismatch; and digesting uncapped dsDNA. Another exemplary method includes synthesizing a first proofread double stranded DNA (dsDNA); synthesizing a second proofread dsDNA; and ligating the first proofread DNA with the second proofread DNA to form a long polynucleotide.

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