title: 18S MiSeq sequencing of AM fungal community
design: Molecular diversity of AM fungi from plant DNA was assessed by 18S rRNA gene amplification with the primers NS31 and AML2 (Simon et al. 1992; Lee et al. 2008) according to Davison et al. (2012). After PCR, the amplification products (pools of 2 PCRs) were purified by using illustra GFX PCR DNA and Gel Band Purification Kit (GE Healthcare Life Sciences, Velizy-Villacoublay, France) following manufacturers guidelines. Then, DNA concentration of PCR products were quantified by using a Qubit fluorometer (Qubit Fluorometric quantitation, Invitrogen) and the Qubit dsDNA HS Assay Kit. PCR product concentration was adjusted to 10ng/l and subjected to paired-end Illumina MiSeq sequencing (2300 bp) by Molecular Research LP (MR DNA, TX, USA). MiSeq Illumina sequencing data were analysed by using Mothur software according the standard operating procedure (http://www.mothur.org/wiki/MiSeq_SOP) proposed in Kozich et al (2013), except that only forward reads were analysed because of the length of PCR products not suitable for paired reads and of a higher quality of forwards reads compared to reverse reads.