An automated multiplex oligonucleotide synthesizer: Development of high-throughput, low-cost DNA synthesis
- 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, Vol. 92, Issue 17; Other Information: PBD: 15 Aug 1995
- Country of Publication:
- United States
- Language:
- English
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