Effects of produced water from oil drilling on soil microbiome using Nanopore Technologies Sequencing
Microbiome DNA was isolated from soil samples using the the DNeasy PowerSoil Pro Kit (Qiagen) according to manufacturers instructions. After isolation DNA concentration and quality was quantified using a Qubit Flex Fluorometer, Nanodrop 2000c Spectrophotometer, and an Agilent Bioanalyzer System. 16S rDNA was amplified using the 16S Barcoding Kit 1-24 SQK-16S024 (Oxford Nanopore Technologies (ONT)). The DNA was amplified by PCR using specific 16S primers (27F and 1492R) that contain 5 tags which facilitate the ligase-free attachment of Rapid Sequencing Adapters. The amplified DNA was sequenced using a R9.4.1 flow cell (FLO-MIN106) on the Oxford Nanopore Technologies MinION sequener producing fastq files.