ena-STUDY-Cairo University-27-12-2017-07:08:15:193-520
fungal metagemoics of the rhizosphere of Rhazya stricta, Enneapogon desvauxii, Citrullus colocynthis, Senna italica, Zygophyllum simplex and the mycoflora of Locust migratoria
The study regionThe site chosen for study was a Rhazya stricta community and public site (Hadda, Mecca-Jeddah road). Sampling site was assigned coordinate via GPS for ease of relocation (N21° 45' 04.03", E39° 53' 88.92").  Field sampling was taken from the rhizosphere of five plants (1A, Rhazya stricta; 2A, Enneapogon desvauxii; 3A, Citrullus colocynthis; 4A, Senna italica; soil5, Zygophyllum simplex), didn't involve plant material from endangered or protected species. Soil cores were taken at each site (2 cm diameter and 10 cm deep), and the number of soil samples collected for each sampling site depending on the heterogeneity of the site and pooled to provide the location sample. Soils were stored in 50 ml Falcons at -20ºC till analysis. In addition, the gut of Locusta migratoria (LocustDNA). DNA Extraction, library construction, and PCR The rules for next-generation sequencing described by Nilsson et al. [27] were followed. The genomic DNA was extracted from 5 g of soil subsample after mixing the various samples of each site by MoBio Power soil kit was done. Once the DNA samples were obtained, a quality test has been done first, then all the qualified DNA is used to construct libraries. For PCR product, the jagged ends of DNA fragment would be converted into blunt ends by using T4 DNA polymerase, Klenow Fragment, and T4 Polynucleotide Kinase. Then add an 'A' base to each 3' end to make it easier to add adapters. After all that, fragments too short would be removed by Ampure beads. For genomic DNA, we use fusion primer with dual index and adapters for PCR, fragments too short would be removed by Ampure beads too. In both cases, only the qualified library can be used for sequencing. Illumina HiSeq/MiSeq encoded amplicons was used for monitoring the diversity of fungi in the rhizosphere of the five plant and Locust's gut. The amplification of the qualified DNA was performed using ITS1 and ITS4 primers as described by Moussa et al. (2017).
The study regionThe site chosen for study was a Rhazya stricta community and public site (Hadda, Mecca-Jeddah road). Sampling site was assigned coordinate via GPS for ease of relocation (N21° 45' 04.03", E39° 53' 88.92").  Field sampling was taken from the rhizosphere of five plants (1A, Rhazya stricta; 2A, Enneapogon desvauxii; 3A, Citrullus colocynthis; 4A, Senna italica; soil5, Zygophyllum simplex), didn't involve plant material from endangered or protected species. Soil cores were taken at each site (2 cm diameter and 10 cm deep), and the number of soil samples collected for each sampling site depending on the heterogeneity of the site and pooled to provide the location sample. Soils were stored in 50 ml Falcons at -20ºC till analysis. In addition, the gut of Locusta migratoria (LocustDNA). DNA Extraction, library construction, and PCR The rules for next-generation sequencing described by Nilsson et al. [27] were followed. The genomic DNA was extracted from 5 g of soil subsample after mixing the various samples of each site by MoBio Power soil kit was done. Once the DNA samples were obtained, a quality test has been done first, then all the qualified DNA is used to construct libraries. For PCR product, the jagged ends of DNA fragment would be converted into blunt ends by using T4 DNA polymerase, Klenow Fragment, and T4 Polynucleotide Kinase. Then add an 'A' base to each 3' end to make it easier to add adapters. After all that, fragments too short would be removed by Ampure beads. For genomic DNA, we use fusion primer with dual index and adapters for PCR, fragments too short would be removed by Ampure beads too. In both cases, only the qualified library can be used for sequencing. Illumina HiSeq/MiSeq encoded amplicons was used for monitoring the diversity of fungi in the rhizosphere of the five plant and Locust's gut. The amplification of the qualified DNA was performed using ITS1 and ITS4 primers as described by Moussa et al. (2017).