RecJ 5' Exonuclease Digestion of Oligonucleotide Failure Strands: A "Green" Method of Trityl-On Purification.

ABSTRACT: Methods of error filtration and correction post-gene assembly are a major bottleneck in the synthetic biology pipeline. Current oligonucleotide purification strategies, including polyacrylamide gel electrophoresis and high-performance liquid chromatography, are often expensive and labor-intensive, give low mass recovery, and contain hazardous chemicals. To circumvent these limitations, we explored an enzymatic means of oligonucleotide purification using RecJ, which is the only known exonuclease to digest single-stranded DNA (ssDNA) in the 5' to 3' direction. As a potential application to remove failure strands generated in oligonucleotide synthesis, we found RecJ does not recognize the 5' dimethoxytrityl blocking group and could therefore be used to specifically target and digest unblocked

SUBMITTER: Jensen M 

PROVIDER: S-EPMC8045232 | biostudies-literature | 2017 May

REPOSITORIES: biostudies-literature