NEXT GENERATION 1,536-WELL OLIGONUCLEOTIDE SYNTHESIZER WITH ON-THE-FLY DISPENSE.

Here we report the development of our Next Generation Automated Multiplexed Oligonucleotide Synthesizer (NG-1536-AMOS), capable of producing 1,536 samples in a single run using a multi-well filtered titer plate. With the potential to synthesize up to 3,456 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 ≤ 5μl with plate speeds up to 150mm/second. Using gas-phase cleavage and deprotection, a full plate of 1,536 60mers can be processed with same-day turnaround with an average yield per well at 3.5 nanomoles. 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 error rates on the NG-1536-AMOS platform at 1.53/717 bases; furthermore, mass spectral analysis on strands synthesized up to 80 bases showed high purity with an average coupling efficiency of 99.5%.