作者
Carles Castaño, Anna Berlin, Mikael Brandström Durling, Katharina Ihrmark, Björn D Lindahl, Jan Stenlid, Karina E Clemmensen, Åke Olson
发表日期
2020/11
期刊
New Phytologist
卷号
228
期号
3
页码范围
1149-1158
简介
- Recent studies have questioned the use of high‐throughput sequencing of the nuclear ribosomal internal transcribed spacer (ITS) region to derive a semi‐quantitative representation of fungal community composition. However, comprehensive studies that quantify biases occurring during PCR and sequencing of ITS amplicons are still lacking.
- We used artificially assembled communities consisting of 10 ITS‐like fragments of varying lengths and guanine‐cytosine (GC) contents to evaluate and quantify biases during PCR and sequencing with Illumina MiSeq, PacBio RS II and PacBio Sequel I technologies.
- Fragment length variation was the main source of bias in observed community composition relative to the template, with longer fragments generally being under‐represented for all sequencing platforms. This bias was three times higher for Illumina MiSeq than for PacBio RS II and Sequel I. All 10 fragments in …
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