作者
Leho Tedersoo, Ave Tooming‐Klunderud, Sten Anslan
发表日期
2018/2
期刊
New Phytologist
卷号
217
期号
3
页码范围
1370-1385
简介
- Second‐generation, high‐throughput sequencing methods have greatly improved our understanding of the ecology of soil microorganisms, yet the short barcodes (< 500 bp) provide limited taxonomic and phylogenetic information for species discrimination and taxonomic assignment.
- Here, we utilized the third‐generation Pacific Biosciences (PacBio) RSII and Sequel instruments to evaluate the suitability of full‐length internal transcribed spacer (ITS) barcodes and longer rRNA gene amplicons for metabarcoding Fungi, Oomycetes and other eukaryotes in soil samples.
- Metabarcoding revealed multiple errors and biases: Taq polymerase substitution errors and mis‐incorporating indels in sequencing homopolymers constitute major errors; sequence length biases occur during PCR, library preparation, loading to the sequencing instrument and quality filtering; primer–template mismatches bias the taxonomic …
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L Tedersoo, A Tooming‐Klunderud, S Anslan - New Phytologist, 2018