Soil.DNA.Replicate3_SamplingTime1_CWF-Treatment
Soil samples of 400mg were used for total genomic DNA extraction using the Nucleospin@ soil kit (Macherey-Nagel). The V3-V4 region of the 16S rDNA, and the fungal ITS1 fungal were then amplified in a PCR reaction using the 341F/805R primer pair for the 16S and ITS1F/58A2R for the ITS, all of which containing the Illumina overhanging adapters. A limited cycle PCR followed in order to attach sample-specific indexes. The amplicons were purified using the Nucleomag@ NGS clean-up and size select kit (Macherey-Nagel). Amplicons were pooled at equimolar concentration and sent for 2x300bp paired-end sequencing using the Illumina Miseq platform. The resulting sequences were de-replicated and quality checked, using the Mothur and Qiime softwares.