metagenomic amplicon sequencing of gene cassettes from polluted river sediment
Total genomic DNA was extracted from individual frozen sediment samples using the PowerSoil DNA isolation kit (MoBio, Carlsbad, CA) according to the manufacturer’s instructions. DNA from sample was amplified using three sets of primer pairs and then the products were purified using a PCR purification kit (Qiagen) and quantified using the Qubit® Fluorometer. afterwards, the Amplicons were then sent for sequencing.