Prunus serotina root metagenome20
We used five nanograms of purified DNA for subsequent metabarcoding of the AMV4.5NF (5-AAGCTCGTAG-TTGAATTTCG-3)/AMDGR (5-CCCAACTATCCCTATTAATCAT-3; Sato et al., 2005) and ITS3_KYO2 (5-GATGAAGAACGYAGYRAA-3)/ITS4 (5-TCCTCCGCTTATTGATATGC-3; Toju et al., 2012; Li et al., 2021) markers, which target the SSU (18S) and ITS2 regions and are usually used to characterize the AMF and ECM communities, respectively. These primers were concatenated to standard Illumina adapters, compatible with Illumina Nextera XT index, which was used to generate the libraries during the sample multiplexing. Purified libraries were pooled at equal molarity, denatured with freshly prepared 0.2 N NaOH, diluted to 4 pM with the pre-chilled hybridization buffer, spiked with a premade PhiX control library at 5% (PhiX control v2, Illumina, USA) and loaded into a MiSeq v3 Reagent Kit (600 Cycles PE, Illumina, USA) to be sequenced in a MiSeq sequencer