丹麦奥尔堡大学Mads Albertsen研究团队近日取得一项新成果。经过不懈努力，他们使用Nanopore或PacBio测序的独特分子标识技术完成了高精度长读扩增子测序。 

在本研究中，研究人员开发了一种高通量扩增子测序方法，该方法结合了独特分子标识（UMI）与牛津纳米孔技术（ONT）或Pacific Biosciences环形共有序列，可获得宏基因组区域内高精度单分子共有序列。研究人员将该方法应用于模式微生物群落核糖体RNA操纵子的扩增子（?4,500 bp）和基因组序列（> 10,000 bp）中，观察到嵌合率<0.02％。要达到平均UMI共识错误率<0.01％，需要UMI读取覆盖率为15倍（ONT R10.3）、25倍（ONT R9.4.1）和3倍（Pacific Biosciences环形共识测序）， 其对应的平均错误率为0.0042％、0.0041％和0.0007％。

据悉，相比于短读测序技术，针对宏基因组区域的高通量扩增子测序仍具有挑战性。

附：英文原文

Title: High-accuracy long-read amplicon sequences using unique molecular identifiers with Nanopore or PacBio sequencing

Author: Sren M. Karst, Ryan M. Ziels, Rasmus H. Kirkegaard, Emil A. Srensen, Daniel McDonald, Qiyun Zhu, Rob Knight, Mads Albertsen

Issue&Volume: 2021-01-11

Abstract: High-throughput amplicon sequencing of large genomic regions remains challenging for short-read technologies. Here, we report a high-throughput amplicon sequencing approach combining unique molecular identifiers (UMIs) with Oxford Nanopore Technologies (ONT) or Pacific Biosciences circular consensus sequencing, yielding high-accuracy single-molecule consensus sequences of large genomic regions. We applied our approach to sequence ribosomal RNA operon amplicons (~4,500bp) and genomic sequences (>10,000bp) of reference microbial communities in which we observed a chimera rate <0.02%. To reach a mean UMI consensus error rate <0.01%, a UMI read coverage of 15× (ONT R10.3), 25× (ONT R9.4.1) and 3× (Pacific Biosciences circular consensus sequencing) is needed, which provides a mean error rate of 0.0042%, 0.0041% and 0.0007%, respectively.

DOI: 10.1038/s41592-020-01041-y

Source: https://www.nature.com/articles/s41592-020-01041-y