Purification of multiplex oligonucleotide libraries by synthesis and selection
Abstract
Complex oligonucleotide (oligo) libraries are essential materials for diverse applications in synthetic biology, pharmaceutical production, nanotechnology and DNA-based data storage. However, the error rates in synthesizing complex oligo libraries can be substantial, leading to increment in cost and labor for the applications. As most synthesis errors arise from faulty insertions and deletions, we developed a length-based method with single-base resolution for purification of complex libraries containing oligos of identical or different lengths. Our method-purification of multiplex oligonucleotide libraries by synthesis and selection-can be performed either step-by-step manually or using a next-generation sequencer. When applied to a digital data-encoded library containing oligos of identical length, the method increased the purity of full-length oligos from 83% to 97%. We also show that libraries encoding the complementarity-determining region H3 with three different lengths (with an empirically achieved diversity >10 6) can be simultaneously purified in one pot, increasing the in-frame oligo fraction from 49.6% to 83.5%.
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Enzymatic DNA synthesis is expected to become both cheaper and faster than phosphoramidite synthesis for data storage applications.
“this approach is expected to become both cheaper and faster than phosphoramidite synthesis for data storage applications.40”
From Emerging Approaches to DNA Data Storage: Challenges and Prospects (2022) · cites this paper as [40]
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Emerging Approaches to DNA Data Storage: Challenges and Prospects
Doricchi, A., Platnich, C. M., Gimpel, A., Horn, F., Earle, M., Lanzavecchia, G., Cortajarena, A. L., Liz-Marzán, L. M., Liu, N., Heckel, R., et al. (2022) · ACS Nano · doi:10.1021/acsnano.2c06748 · Cited by 235 · see citing works
- Authors
- Choi, H., Choi, Y., Choi, J., Lee, A. C., Yeom, H., Hyun, J., Ryu, T., & Kwon, S.
- Affiliations
- Seoul National University (KR)
- Harvard University (US)
- Corresponding author
- Ryu, Taehoon
- Kwon, Sunghoon
- Venue
- Nature Biotechnology
- Published
- 2021-07-29
- Volume
- 40
- Issue
- 1
- Pages
- 47-53
- ISSN
- 0733-222X (ISSN-L)
- 1087-0156
- 1546-1696
- OpenAlex
- W3183621268
- Cited by
- 32
- References
- 56
- Citations by year
- 2026: 5 · 2025: 10 · 2024: 3 · 2023: 2 · 2022: 9 · 2021: 3
- FWCI
- 1.33
- Type
- journal article
- Language
- en
- Open access
- closed
- Licence
- Locations
- Nature Biotechnology (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Research Foundation of Korea (2020M3H1A1073304, 2021R1C1C2010079, 2018M3A9D7079488, 2020R1A3B3079653, 2015K1A4A3047345)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA and Nucleic Acid Chemistry
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA and Biological Computing
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA and protein synthesis mechanisms
- Keywords
- insertion and deletion errors, single-base resolution, next-generation sequencing, library diversity
- MeSH terms
- DNA, Oligonucleotides
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