期刊
ENVIRONMENTAL MICROBIOLOGY REPORTS
卷 11, 期 5, 页码 659-668出版社
WILEY
DOI: 10.1111/1758-2229.12776
关键词
-
资金
- Estonian Science Foundation, RMK [PUT1399P, MOBERC13]
Development of high-throughput sequencing techniques has greatly benefited our understanding about microbial ecology, yet the methods producing short reads suffer from species-level resolution and uncertainty of identification. Here, we optimize Pacific Biosciences-based metabarcoding protocols covering the internal transcribed spacer (ITS region) and partial small subunit of the rRNA gene for species-level identification of all eukaryotes, with a specific focus on Fungi (including Glomeromycota) and Stramenopila (particularly Oomycota). Based on tests on composite soil samples and mock communities, we propose best suitable degenerate primers, ITS9munngs + ITS4ngsUni for eukaryotes and selected groups therein and discuss the pros and cons of long read-based identification of eukaryotes.
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