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Democratizing water monitoring: Implementation of a community-based qPCR monitoring program for recreational water hazards

biorxiv. 2020; 
Sydney P Rudko, Ronald R Reimink, Bradley Peter, Jay White, Patrick C Hanington
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Gene Synthesis Samples were quantitated relative to a plasmid standard curve which contained 50,000, 5000, 500, 50, 5 and 0.5 copies. Each of the gene targets below was synthesized (IDTDNA) into a puc19 plasmid vector (Genscript). Thermocycling was performed on the ABI 7500 Fast or the QuantStudio 3 using a standard, 40 cycle, two-step reaction. The thermocycling parameters were a 30 second hold at 95 degrees, followed by a 30 second denaturation cycle at 95 degrees, and a 60 degrees annealing cycle. Each qPCR reaction had a final volume of 20, and we added 5μL of DNA to each reaction. Get A Quote

摘要

Recreational water monitoring can be challenging due to the highly variable nature of pathogens and indicator concentrations, the myriad of potential biological hazards to measure for, and numerous access points, both official and unofficial, that are used for recreation. The aim of this study was to develop, deploy, and assess the effectiveness of a quantitative polymerase chain reaction (qPCR) community-based monitoring (CBM) program for the assessment of bacterial and parasitic hazards in recreational water. This study developed methodologies for performing qPCR ‘in the field’, then engaged with water management and monitoring groups, and tested the method in a real-world implementation study to evaluate... More

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