Next generation 1536-well oligonucleotide synthesizer with on-the-fly dispense

作者：Jensen Michael^*; Roberts Lester; Johnson Andrew; Fukushima Marilyn; Davis Ronald

来源：Journal of Biotechnology, 2014, 171: 76-81.

DOI：10.1016/j.jbiotec.2013.11.027

• Oligonucleotide
• Synthesis automation
• Chemical DNA synthesis
• Synthetic biology

Here we report the development of our Next Generation Automated Multiplexed Oligonucleotide Synthesizer (NG-1536-AMOS), capable of producing 1536 samples in a single run using a multi-well filtered titer plate. With the potential to synthesize up to 3456 samples per plate, we converted the BioRAPTR Flying Reagent Dispenser into an open-well system where spent reagents are drained to waste under vacuum. During synthesis, reagents are delivered on-the-fly to each micro-titer well at volumes <5 pi with plate speeds up to 150 mm/s. Using gas-phase cleavage and deprotection, a full plate of 1536 60mers may be processed with same-day turnaround with an average yield per well at 3.5 nmol. Final product at only $0.00277/base is eluted into a low-volume collection plate for immediate use in downstream application (e.g. Biomek FX for versatile sample handling). Also, crude oligonucleotide quality is comparable to that of commercial synthesis instrumentation, with an error rate on the NG-1536-AMOS platform of 1.531717 bases. Furthermore, mass spectral analysis on strands synthesized up to 80 bases showed high purity with an average coupling efficiency of 99.5%.

• 出版日期2014-2-10