Titel
Chemical and photochemical error rates in light-directed synthesis of complex DNA libraries
Autor*in
Adrien Leger
European Molecular Biology Laboratory, European Bioinformatics Institute, Wellcome Genome Campus
Autor*in
Yaniv Erlich
Erlich Lab LLC, Raanana, Israel
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Abstract
Nucleic acid microarrays are the only tools that can supply very large oligonucleotide libraries, cornerstones of the nascent fields of de novo gene assembly and DNA data storage. Although the chemical synthesis of oligonucleotides is highly developed and robust, it is not error free, requiring the design of methods that can correct or compensate for errors, or select for high-fidelity oligomers. However, outside the realm of array manufacturers, little is known about the sources of errors and their extent. In this study, we look at the error rate of DNA libraries synthesized by photolithography and dissect the proportion of deletion, insertion and substitution errors. We find that the deletion rate is governed by the photolysis yield. We identify the most important substitution error and correlate it to phosphoramidite coupling. Besides synthetic failures originating from the coupling cycle, we uncover the role of imperfections and limitations related to optics, highlight the importance of absorbing UV light to avoid internal reflections and chart the dependence of error rate on both position on the array and position within individual oligonucleotides. Being able to precisely quantify all types of errors will allow for optimal choice of fabrication parameters and array design.
Stichwort
Genetics
Objekt-Typ
Sprache
Englisch [eng]
Persistent identifier
https://phaidra.univie.ac.at/o:1539058
Erschienen in
Titel
Nucleic Acids Research
Band
49
Ausgabe
12
ISSN
0305-1048
Erscheinungsdatum
2021
Seitenanfang
6687
Seitenende
6701
Verlag
Oxford University Press (OUP)
Projektnummer
P27275 – Austrian Science Fund (FWF)
Projektnummer
P30596 – Austrian Science Fund (FWF)
Erscheinungsdatum
2021
Zugänglichkeit
Rechteangabe
© The Author(s) 2021

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