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One Step Growth Curve Of T4 Bacteriophage In E.coli

At this time, phage particles begin to accumulate in the extracellular environment (Flint 2000). The rise period is followed by the fifth and final plateau period when the accumulation of phage particles ceases. This plateau can be seen as the latent period of the following multiplication cycle (Flint, 2000). .
             One benefit of the one step growth curve is that it allows the calculation of burst size (Flint, 2000). Burst size is the number of mature phage particles produced during the multiplication round. Calculation of burst size is one of the major goals in a one step growth curve experiment (Flint, 2000). .
             The goal of this experiment is to use the one step growth curve method to visualize and describe the T4 phage lytic cycle in Escherichia coli. In addition, the burst size for T4 bacteriophage will be obtained from growth curve data.
             Methods.
             The host organism used in this experiment was log phase E.coli at a stock concentration of 2x108 cells per milliliter. At the beginning of the experiment (time = 0 minutes) 100L of the E.coli was mixed with 100 L of T4 phage. The T4 phage used was at a concentration of 10x108 PFU per milliliter. This produced an MOI ratio of five phage for every cell. This cell/phage mixture was labeled ADS and incubated in a heating block for five minutes at 37C. .
             At the five-minute mark, the first student diluted 10L of the ADS 100 fold using sterile broth as the diluent and placed the diluted ADS into a tube labeled UP 10-2. The UP tube determined the amount of unadsorbed phage in the sample. The UP10-2 tube contained 3 drops of chloroform. The UP 10-2 tube was mixed and then left to settle. .
             Three one hundred fold dilutions were then made from 10L of the ADS tube using sterile broth as a diluent. These dilutions were placed in tubes labeled IC 10-2, IC 10-4 and IC 10-6 respectively. The IC tubes were used to determine the total infectious centers in the ADS tube.


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