Environmental sample from arable field
AM fungal sequences were amplified from soil DNA extracts using AM fungal-specific primers for the smallsubunit (SSU) ribosomal RNA gene: WANDA and AML2. A first PCR was conducted with amplicon specific primers linked to Illumina Nextera XT sequencing adapters (Illumina forward primer adaptor: 5TCGTCGGCAGCGTCAGATGTGTATAAGAGACAG3; Illumina reverse primer adaptor: 5GTCTCGTGGGCTCGGAGATGTGTATAAGAGACAG3). The reaction mix contained 12.5 l KAPA HiFi Hotstart PCR mix ; 1.0 l of each 10 M primer; 6 l of template DNA; and MQ water to reach a total reaction volume of 25 l. The PCR was performed under the following cycling conditions: 95C for 3 min, 40 cycles of 95C for 30 s, 55C for 30 s, 72C for 30 s followed by 72C for 5 min. A second PCR was performed with Nextera XT indexadapters. The reaction mix contained 15 l KAPA HiFi Hotstart PCR mix; 5 l of Nextera XT index 1 Primer (N7xxx) and 5 l of Nextera XT index 2 Primer (E5xxx); 5 l of DNA (10 ng l1), and MQ water to reach a total reaction volume of 30 l. PCR was performed under the following cycling conditions: 95C for 3 min, seven cycles of 95C for 30 s, 55C for 30 s, 72C for 30 s, followed by 72C for 5 min. After the second PCR, samples were purified with Agencourt AMPure XP beads. The resulting mix was sequenced on the Illumina MiSeq platform, using a 2 x 300 bp pairedend sequencing approach.