title: Wheat and couchgrass rhizosphere microbiome
design: Total rhizospheric DNA of wheat and couchgrass samples was extracted using the FastDNA SPIN Kit for Soil (MP Biomedicals, USA) according to the manufacturer's instructions. The 16S rRNA gene was amplified using universal bacterial primers: 27f: 5-AGA GTT TGA TCC TGG CTC AG-3 and 1492r: 5-ACG GYT ACC TTG TTA CGA CTT-3. The 25 L PCR reaction mixture contained 1 L of DNA template, 0.5 L each of forward and reverse primers (10 M), 2.5 L dNTPs (10 M), 2.5 L of 10x buffer with SYBR Green, 2.5 L MgCl2 (25 M), 0.2 L SynTaq polymerase (5 U L1), and 15.3 L of nuclease-free water. Thermocycling was performed as follows: initial denaturation at 94 for 5 min, 35 three-step cycles at 94 for 1 min, at 55 for 1 min and at 72 for 2 min, followed by a final elongation step at 72 for 10 min. Sequencing was conducted on the Illumina NextSeq 500 platform at the Interdisciplinary Center for Proteomic Research, Kazan Federal University (Kazan, Russia). Sequence data were analyzed using QIIME version 1.6.0 (East Lansing, MI, USA). The taxonomic classification was assigned using the Silva 138 database.
library_name: Wheat_rhizosphere_metagenome35
identifiers.namespace: SUB15554923
identifiers.id: Wheat_rhizosphere_metagenome35