The Escherichia coli species is a human fecal contamination indicator and as such is used in beach monitoring efforts. Quantifying E. Coli presence in local beach waters helps the health department determine if a beach should be closed. The current method of determination, Colilert, takes 18 hours to produce data. Quantitative Polymerase Chain Reaction (qPCR), which measure genetic DNA, is also method used to quantify the number of E. Coli, but it can be done much faster than Colilert. In order to standardize the qPCR results, an internal standard is included which is salmon DNA. This project goes through the process of optimizing the salmon standard curve. Each of the components was modified and the resulting standard curve was analyzed for improvements; the primers and probe were purchased new and the concentrations were varied, the DNA was purchased new and the standard curve concentrations and dilution methods were varied, the DNA was cleaned with a Qiagen kit, and new master mix and bovine serum albumin were purchased and prepared. We found that changes to the concentrations of primers and probe and cleaning the DNA showed an increase of optimization, and that changing the dilution methods had no effect of optimization. A combination of the above modifications may be able to produce an optimized salmon DNA standard curve.
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