An automated multiplex oligonucleotide synthesizer: Development of high-throughput, low-cost DNA synthesis

Journal Article · Tue Aug 15 04:00:00 EDT 1995 · Proceedings of the National Academy of Sciences of the United States of America

DOI:https://doi.org/10.1073/pnas.92.17.7912· OSTI ID:98861

Lashkari, D A; Hunicke-Smith, S P; Norgren, R M ^[1]

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1. Stanford Univ. School of Medicine, Stanford, CA (United States); and others

An automated oligonucleotide synthesizer has been developed that can simultaneously and rapidly synthesize up to 96 different oligonucleotides in a 96-well microtiter format using phosphoramidite synthesis chemistry. A modified 96-well plate is positioned under reagent valve banks, and appropriate reagents are delivered into individual wells containing the growing oligonucleotide chain, which is bound to a solid support. Each well has a filter bottom that enables the removal of spent reagents while retaining the solid support matrix. A seal design is employed to control synthesis environment and the entire instrument is automated via computer control. Synthesis cycle times for 96 couplings are <11 min, allowing a plate of 96 20-mers to be synthesized in <5 hr. Oligonucleotide synthesis quality is comparable to commercial machines, with average coupling efficiencies routinely >98% across the entire 96-well plate. No significant well-to-well variations in synthesis quality have been observed in in >6000 oligonucleotides synthesized to date. The reduced reagent usage and increased capacity allow the overall synthesis cost to drop by at least a factor of 10. With the development of this instrument, it is now practical and cost-effective to synthesize thousands to tens of thousands of oligonucleotides. 22 refs., 4 figs., 1 tab.

• Sponsoring Organization: USDOE
• OSTI ID: 98861
• Journal Information: Proceedings of the National Academy of Sciences of the United States of America, Journal Name: Proceedings of the National Academy of Sciences of the United States of America Journal Issue: 17 Vol. 92; ISSN PNASA6; ISSN 0027-8424
• Country of Publication: United States
• Language: English

Related Subjects

55 BIOLOGY AND MEDICINE
BASIC STUDIES
COST ESTIMATION
DNA
DNA SEQUENCERS
OLIGONUCLEOTIDES
SYNTHESIS