title: Amplicon sequences of AMF in soil
design: DNA was isolated from a ~ 0.25g subsample of each soil sample using the FastDNA  SPIN Kit for Soil, in combination with the FastPrep  Instrument (MP Biomedicals, Santa Ana, CA). For amplification, DNA was diluted to 5 ng/ l. We used the primer set FLR3 / FLR4 to amplify the D2 region of the LSU rDNA. Individual Golay barcodes were attached to the reverse primer of each sample. We used the Phusion High-Fidelity PCR Master Mix with HF Buffer with the following thermocycling parameters: Initial denaturation 1 min at 95_C, 36 cycles of 1 min at 95_C (denaturation), 1 min at 57_C (annealing), and 90 s at 72_C (extension), with a final extension phase of 5 min at 72_C. For each sample, triplicate PCR products were obtained and composited. Amplified DNA was extracted from 1.5 % low EEO agarose gels and cleaned with the Promega Wizard SV Gel and PCR Clean-Up System. Subsequently DNA concentrations were determined (Image J   Image Processing and Analysis in Java, and Nanodrop (Nanodrop  ND-1000). Purified DNA of all 20 samples were pooled in equimolar amounts.