title: metagenome of terrestrial vertebrates:soil
design: DNA extraction was carried out using the DNeasy PowerLyzer PowerSoil Kit (QIAGEN) automated on the QIAGEN QIAcube (QIAGEN). The preparation of the DNA library was performed using a one-step PCR method. PCR primers designed by Riaz et al. (2011), targeting the V5 region of 12s ribosomal RNA (rRNA), were modified to include the addition of Illumina sequencing adapters and unique dual index combinations. PCR products were amplified using the Platinum Taq DNA Polymerase High Fidelity kit (Invitrogen). Reactions were assembled using an epMotion 5075 Liquid Handler (Eppendorf). The library was sequenced using a 300 cycle Illumina MiSeq kit, using custom sequencing and index primers.  RIAZ, T., SHEHZAD, W., VIARI, A., POMPANON, F., TABERLET, P. & COISSAC, E. 2011. ecoPrimers: inference of new DNA barcode markers from whole genome sequence analysis. Nucleic acids research, 39, e145-e145.