Pearl millet soil metabarcoding
Total RNA was extracted from 0.4 g of roots, and 2 g of root-adhering soil or bulk soil using the RNeasy PowerSoil Total RNA Kit, Qiagen. DNA digestion was performed using the TURBO DNA-freeTM Kit (Invitrogen) followed by RNA purification using the RNeasy Mini Kit (Qiagen). cDNA was synthesized using the Transcriptor First Strand cDNA synthesis Ki, V.6 (Roche) and were used for PCR targeting bacteria and fungi, using V4 and V3-V4 16S rRNA, and fITS7 and ITS4 respectively. The primers were designed to contain overhang sequences compatible with Illumina Nextera XT index. The purified amplicons were sequenced by Biofidal Laboratory (Lyon, France) using the MiSeq, Illumina platform.