Beilstein J. Org. Chem.2023,19, 1957–1965, doi:10.3762/bjoc.19.146
have difficulty to produce sequences containing stable secondary structures. Here, we report a direct de novo chemical synthesis of 400 nt ODNs, and their isolation from the complex reaction mixture using the catching-by-polymerization (CBP) method. To determine the authenticity of the ODNs, 399 and
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Keywords: automated synthesis; catching-by-polymerization; gene assembly; long oligonucleotide; synthetic biology; Introduction
Long oligodeoxynucleotides (ODNs) are segments of DNAs extending beyond one hundred nucleotides (nt). Emerging research areas such as synthetic biology [1][2], protein
impossible [20]. Once the length of ODNs reaches about 100-mer, RP HPLC could not resolve the full-length and failure sequences. Therefore, to isolate the full-length sequence, the catching-by-polymerization (CBP) method was employed (step 2, Figure 1, and Scheme 1). To the crude ODN, the polymerization
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Graphical Abstract
Figure 1:
Workflow for the construction, verification, and expression of the 800 bp GFP gene from chemically ...