High throughput parallel synthesis of oligonucleotides with 1536 channel synthesizer.
A 1536 channel oligonucleotide synthesizer, the MultiSyn, was developed with the capability to simultaneously synthesize 1536 oligonucleotides of 20mer length in 10 h. The instrument was designed to synthesize different sequences of various lengths in micro-wells and has synthesized oligonucleotides as long as 119 nt with reasonably good yields using CPG beads of 1000 A pore size. The instrument consists of four 384 channel synthesis modules. Phosphoramidite chemistry was employed and step yields as high as 99.3% were achieved. The enhancement of oligonucleotide synthesis throughput is accomplished by increasing the spatial density of reaction wells. We have identified several parameters that are critical in achieving a good synthesis yield and negligible failure rate in small reaction wells. The coefficient of variation (CV) of product yields in 1536 reaction wells was 20%. The quality of the product was examined by capillary electrophoresis and mass spectrometry. The instrument has robustly synthesized oligonucleotides of various lengths for use as primers and probes for PCR amplifications, oligonucleotide microarrays and genotyping applications. This high throughput oligonucleotide synthesizer is a useful instrument for genomic applications, which require tens of thousands of probes or primers in a short time.
Cheng
Ji-Yen
JY
Institute of Biomedical Sciences, Academia Sinica, Taipei, Taiwan 115, Republic of China.
Chen
Hung-Hui
HH
Kao
Yu-Shu
YS
Kao
Wei-Chen
WC
Peck
Konan
K
D001483:Base Sequence; D002625:Chemistry, Organic / Q000295:instrumentation* / Q000379:methods; D004247:DNA / Q000138:chemical synthesis* / Q000737:chemistry; D019075:Electrophoresis, Capillary; D013058:Mass Spectrometry; D009841:Oligonucleotides / Q000138:chemical synthesis* / Q000737:chemistry; D015203:Reproducibility of Results
D009841:Oligonucleotides; D004247:DNA