Effects of produced water from oil drilling on pregnant mice and their pups microbiome across multiple gestation periods using Nanopore Technologies Sequencing
Caecum samples were removed from the mice post mortum, both the dams and their pups (the pups were never introduced to the produced water except through their mothers). Microbiome DNA was isolated from the whole caecum samples (only the 0% and 100% for the dams and 0%, 50%, and 100% for the pups) using the Qiagen DNeasy PowerSoil Pro Kit according to the manufacturer's 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 Oxford Nanopore Technologies 16S Barcoding Kit. The DNA was amplified by PCR using specific 16S primers, 27F adn 1492R. The amplified DNA was sequenced using a R9.4.1 flow cell on the Oxford Nanopore Technologies MinION sequencer.