High Throughput Multiplex Oligonucleotide Synthesizer

10.30047/JGMB.200006.0012

Ji-Yen Cheng；Konan Peck

Journal of Genetics and Molecular Biology

11卷2期（2000 / 06 / 01）

86 - 86

英文

The demand for oligonucleotides (probes or PCR primers) will soon rise dramatically when the postgenome era comes, in large-scale application such as single nucleotide polymorphism (cSNP) study, artificial gene construction, and gene amplification directly from the open reading frame. However, purchasing a large number of primers from commercial sources costs a lot. In addition, the synthesis throughput is too low to meet the need of large-scale studies. We have constructed a multiplex oligonucleotide synthesizer capable of producing 96 20-mers in 5 hours. For each oligonucleotide, 20 nmol is prepared. Standard phosphoramidite chemistry is used. The instrument reduces the reagent cost of each base to ~1/10 of commercial price. Additionally, by introducing the 384-well plate, the throughput can be easily extended to 4 folds. For comparison, ABI 3948 synthesizer can produce 48 oligonucleotides in 24 hours. The performance and possible application of the instrument is described in this poster. The product purity and quality are evaluated by HPLC and UV-Vis spectroscopy. The biological activity of the primer product has been confirmed by PCR and DNA/DNA hybridization experiment.