Fully Automated Parallel Oligonucleotide Synthesizer

Abstract

Design and construction of automated synthesizers using the tilted plate centrifugation technology is described. Wash solutions and reagents common to all synthesized species are delivered automatically through a 96-channel distributor connected to a gear pump through two four-port selector valves. Building blocks and other specific reagents are delivered automatically through banks of solenoid valves, positioned over the individual wells of the microtiterplate. These instruments have the following capabilities: Parallel solid-phase oligonucleotide synthesis in the wells of polypropylene microtiter plates, which are slightly tilted down towards the center of rotation, thus generating a pocket in each well, in which the solid support is collected during centrifugation, while the liquid is expelled from the wells. Eight microtiterplates are processed simultaneously, providing thus a synthesizer with a capacity of 768 parallel syntheses. The instruments are capable of unattended continuous operation, providing thus a capacity of over two millions 20-mer oligonucleotides in a year.

Substrate Cleavage Analysis of Furin and Related Proprotein Convertases

“…The oligonucleotide tags were synthesized using a highthroughput oligonucleotide synthesizer at Illumina, San Diego, CA (27,31). Oligonucleotides were modified by coupling a phosphoramidite containing a formylindole group (Link Technologies, Bellshill, Scotland) as the last building block in DNA synthesis.…”

Highly Parallel SNP Genotyping

“…SNP assays require one or more oligonucleotides (the GoldenGate assay requires 3n + 3 oligonucleotides for n SNPs), which are most efficiently generated by de novo chemical synthesis. Anticipating a need to create millions of SNP assays, we developed a high-throughput, low-cost centrifugal oligonucleotide synthesizer (Lebl et al 2001). This LIMS-integrated automated instrument is able to produce hundreds to thousands of oligonucleotides per day.…”

A High-Complexity, Multiplexed Solution-Phase Assay for Profiling Protease Activity on Microarrays

“…The tag DNA part of the molecule was synthesized utilizing the high-throughput oligonucleotide synthesizers developed by Illumina [46,52]. Oligonucleotides were modified by attachment of either an amino linker at 5' end of the DNA molecule, followed by condensation of 4-carboxybenzaldehyde, or by coupling a phosphoramidite containing a formylindole group (Link Technologies, Bellshill, Scotland) as the last building block in DNA synthesis.…”