Simultaneous and stoichiometric purification of hundreds of oligonucleotides

Alessandro Pinto, Sherry X. Chen and David Yu Zhang ()
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Alessandro Pinto: Rice University
Sherry X. Chen: Rice University
David Yu Zhang: Rice University

Nature Communications, 2018, vol. 9, issue 1, 1-9

Abstract: Abstract Purification of oligonucleotides has traditionally relied on mobility-based separation methods. However, these are imperfect, biased, and difficult to scale high multiplex. Here, we present a method for simultaneous purification of many oligonucleotides that also normalizes concentrations. The method uses a rationally designed randomer capture probe to enrich for oligos with perfect 5′ sequences, based on the observation that synthesis errors are correlated: product molecules with one or more deletions in one region are also more likely to have deletions in other regions. Next-generation sequencing analysis of 64-plex 70 nt purification products show a median 78% purity, a significant improvement over polyacrylamide gel electrophoresis and high pressure liquid chromatography (60% median purity). Additionally, 89% of the oligo products are within a factor of 2 of the median concentration.

Date: 2018
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DOI: 10.1038/s41467-018-04870-w

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