DNA seq soil microbiome52fungisoilnot fertilizednot inoculatedno trichoyes plantyes soil3
The 16S rRNA gene V4 variable region (for bacteria communities) and ITS1 gene (for fungal communities) were analyzed. Polymerase chain reaction (PCR) primers 515/806 and ITS1F-Bt1/ITS2R, for bacterial and fungal communities respectively, were used in a single-step 30 cycle PCR using the HotStarTaq Plus Master Mix Kit (Qiagen, USA). The PCR conditions were: 94C for 3 minutes, followed by 30 cycles (5 cycle used on PCR products) of 94C for 30 seconds, 53C for 40 seconds and 72C for 1 minute, after which a final elongation step at 72C for 5 minutes was performed. Sequencing was performed at MR DNA (www.mrdnalab.com, Shallowater, TX, USA) on an Ion Torrent (PGM) following the manufacturers guidelines.