Preliminary Work for Low Temperature Phage Amplification

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Abstract:

This project determined the growth curve for BL21 Escherichia coli and compared two propagation methods for T7 bacteriophage to determine which resulted in a higher titer solution of bacteriophage. A high titer working stock solution is desirable to ensure the lysate is stable for future use and development [19]. Both methods were adapted from Dr. Bonillas Phage on Tap protocol [3]. The first method uses a set volume of 1 mL of bacteriophage to a 1 mL solution of E. coli culture. The second uses a volume determined by the multiplicity of infection (MOI), which is the ratio of E. coli colony forming units to the plaque forming units of T7 phage. It was determined that the propagation method using MOIappeared to result in a higher titer working stock solution; however, these results were not statistically different. The p-value obtained was close to the selected alpha-value, so it is believed that with a larger sample size the result should be statistical different. This project is part of a much larger effort. This project lays out the ground work for developing a genetically engineered bacteriophage capable of detecting and removing E. coli from water samples at room temperature (20-22 deg C).

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