小柯机器人

韩国首尔国立大学教授Kwon, Sunghoo和ATG生物技术公司Taehoon Ryu团队合作，通过合成和筛选纯化了多重寡核苷酸库。2021年7月29日出版的《自然-生物技术》发表了这项成果。

由于合成寡核苷酸文库的大多数错误是由错误插入和删除引起的，研究人员开发了一种基于长度的方法，具有单碱基分辨率，以用于纯化含相同或不同长度的寡核苷酸复杂文库。该方法通过合成和选择纯化多重寡核苷酸文库从而可以逐步手动执行，也可以使用下一代测序仪进行。当用于含相同长度的寡核苷酸数据库构建时，该方法将全长寡核苷酸的纯度从83%提高到97%。

研究还表明，可以在一次反应中同时纯化编码三种具有不同互补决定区长度的H3文库（根据经验获得的多样性>106），并将寡核苷酸分数从49.6%增加到 83.5%。

据悉，复杂的寡核苷酸 (oligo) 文库是合成生物学、药物生产、纳米技术和依赖于DNA数据存储等多种应用的基础。然而，合成复杂寡核苷酸文库的错误率比较高，这增加了应用程序的成本和劳动力。

附：英文原文

Title: Purification of multiplex oligonucleotide libraries by synthesis and selection

Author: Choi, Hansol, Choi, Yeongjae, Choi, Jaewon, Lee, Amos Chungwon, Yeom, Huiran, Hyun, Jinwoo, Ryu, Taehoon, Kwon, Sunghoon

Issue&Volume: 2021-07-29

Abstract: Complex oligonucleotide (oligo) libraries are essential materials for diverse applications in synthetic biology, pharmaceutical production, nanotechnology and DNA-based data storage. However, the error rates in synthesizing complex oligo libraries can be substantial, leading to increment in cost and labor for the applications. As most synthesis errors arise from faulty insertions and deletions, we developed a length-based method with single-base resolution for purification of complex libraries containing oligos of identical or different lengths. Our method—purification of multiplex oligonucleotide libraries by synthesis and selection—can be performed either step-by-step manually or using a next-generation sequencer. When applied to a digital data-encoded library containing oligos of identical length, the method increased the purity of full-length oligos from 83% to 97%. We also show that libraries encoding the complementarity-determining region H3 with three different lengths (with an empirically achieved diversity >106) can be simultaneously purified in one pot, increasing the in-frame oligo fraction from 49.6% to 83.5%.

DOI: 10.1038/s41587-021-00988-3

Source: https://www.nature.com/articles/s41587-021-00988-3

Nature Biotechnology：《自然—生物技术》，创刊于1996年。隶属于施普林格·自然出版集团，最新IF：68.164
官方网址：https://www.nature.com/nbt/
投稿链接：https://mts-nbt.nature.com/cgi-bin/main.plex