کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
23246 43424 2014 6 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Next generation 1536-well oligonucleotide synthesizer with on-the-fly dispense
ترجمه فارسی عنوان
نسل بعدی 1539 سینت سایزر الیگونوکلئوتید با استفاده از آن در پرواز
موضوعات مرتبط
مهندسی و علوم پایه مهندسی شیمی بیو مهندسی (مهندسی زیستی)
چکیده انگلیسی


• 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%.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Biotechnology - Volume 171, 10 February 2014, Pages 76–81
نویسندگان
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