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

Paper ID Volume ID Publish Year Pages File Format Full-Text
23246 43424 2014 6 PDF Available
Title
Next generation 1536-well oligonucleotide synthesizer with on-the-fly dispense
Abstract

•On average, 3.5 nmol per well.•No post-synthesis amplification required.•Samples are not heterogeneously pooled.•Oligonucleotide quality at 99.5% coupling efficiency for 80mers.•1.53 errors/717 bases.

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 μl 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 1.53/717 bases. Furthermore, mass spectral analysis on strands synthesized up to 80 bases showed high purity with an average coupling efficiency of 99.5%.

Keywords
Oligonucleotide; Synthesis automation; Chemical DNA synthesis; Synthetic biology
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Next generation 1536-well oligonucleotide synthesizer with on-the-fly dispense
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 171, 10 February 2014, Pages 76–81
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us