High-accuracy long-read amplicon sequences using unique molecular identifiers with Nanopore or PacBio sequencing
High-accuracy long-read amplicon sequences using unique molecular identifiers with Nanopore or PacBio sequencing
复制标题
DOI：
10.1038/s41592-020-01041-y
复制
发表时间：
2021-01-11
期刊：
影响因子：
48
通讯作者：
Albertsen, Mads
中科院分区：
文献类型：
--
作者：
Karst, Soren M.;Ziels, Ryan M.;Albertsen, Mads
大基因组区域的高通量扩增子测序对于短读技术来说仍然具有挑战性。在这里，我们报告了一种高通量扩增子测序方法，将独特分子标识符（UMIs）与牛津纳米孔技术（ONT）或太平洋生物科学循环共识测序相结合，产生高精度的大基因组区域单分子共识序列。我们应用我们的方法对参考微生物群落的核糖体RNA操纵子扩增子（类似于4,500 bp）和基因组序列（>10,000 bp）进行测序，我们观察到嵌合体率
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 (similar to 4,500 bp) and genomic sequences (>10,000 bp) of reference microbial communities in which we observed a chimera rate