title: wastewater effluent sample cDNA derived from Stage 2, sequenced in Illumina
design: Wastewater samples were collected from the various stages of the wastewater treatment process at the Doha West Wastewater Treatment plant. Briefly, the samples were collected in sterilized glass bottles and directly stored in an ice Box of around 4 C until transportation to the laboratory. Wastewater samples were filtered through sterile 0.22 m Durapore hydrophilic PVDF sterile membrane (Millipore, Darmstadt, Germany). Sampling and preservation strategies were conducted according to the APHA 2008. RNA was isolated, according to the manufacturers protocol, from 25 ml of each sample using the RNeasy PowerWater Kit (Qiagen, Hilden, Germany). PCR primers 515F: 5'- GTGCCAGCMGCCGCGGTAA-3' and 806R: 5'-GGACTACHVHHHTWTCTAAT-3' (Integrated DNA Technologies, Iowa, USA) were used to amplify the V4 hypervariable region of the 16SrRNA from the isolated samples. PCR reaction was carried out and paired -end sequencing of the library was done on the Miseq platform based on the standard protocol.