title: LSU metabarcodes:Tomicus piniperda
design: In the laboratory, the specimens were rinsed with pure water to remove loosely adhering fungal tissue, and thoroughly macerated individually to ensure that all fungi associated with the specimens were released. DNA was extracted using the Qiagen DNeasy Blood and Tissue spin column extraction kit (Qiagen, Valencia, CA, USA). The DNA extracts were then used for fungal metabarcoding of LSU using primers LR0R (5-ACCCGCTGAACTTAAGC-3) (Vilgalys and Hester, 1990) / JH-LSU-369rc (5-CTTCCCTTTCAACAATTTCAC-3) (Li et al., 2016) targeting the D1-D2 region at the 5 end of the LSU gene immediately downstream of the ITS2 region.Raw reads were demultiplexed, primer trimmed and singleton reads removed with Cutadapt (Martin, 2011).