title: miseq of Bulk soil: Alley cropping
design: Distribution of rhizosphere fungal communities using sequencing of PCR amplicon of ITS rDNA was conducted with the illumina MiSeq platform (MRDNA Laboratories, Shallowater, TX, United States) targeting the V3-V4 hyper variable region using fungal universal (ITS1/ITS2) where amplicon sequencing (bTEFAP) was performed and used for the fungal analysis. A single-step 30-cycle polymerase chain reaction (PCR with HotStart Taq Plus Master Mix kit (Qiagen, Valencia, CA, United States) was employed. Samples were sequenced utilizing Illumina MiSeq chemistry following the manufacturers protocols. The PCR product were analyzed on 2% agarose gel. Illumina library preparations of the amplicons of the amplified and cleaned PCR products were performes. The library quality was assessed on the Qubil@ 3.0 Fluorometer (Thermo Scientific) and Agilent Bioanalyzer 2100 system.