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Experiment #2 – One Step Growth Curve
Introduction
Viruses (or virulent phages) are particles that are incapable of reproducing on their own without the aid of a suitable host cell to provide cellular machinery. ... It was determined in 1939 by Emory Ellis and Max Delbruck that the single-cell life cycle of viruses could be analyzed by the one-step growth analysis (Flint, et al, 2000, pp 40).
A One Step Growth Curve is a useful tool in determining information about the lytic cycle of a specific virus, however, there are a few things to keep in mind before proceeding to study the single life cycle. The most crucial aspect of the experiment is that the infection of the bacterial cells be synchronized to ensure that the various phases of replication within an infected cell population are essentially happening at the same time, as if it were a single interaction (or as “one step”). To ensure that this does occur, it is important to make sure that each cell be infected by at least one virus particle. ... Once adsorption has occurred, the entire mixture is diluted to prevent further interaction between the bacterial particles and the phage, and it’s this step that effectively synchronizes the various periods of the lytic cycle of the virus. ...
The burst size is actually the yield of infectious virus per infected cell and can be calculated from the overall rise in phage titre from the one-step growth curve. ... Plotting the phage count versus time gives an illustration of the single-step growth response of the T4 after initiating infection of E. ... This step indicated whether any phage was in the original bacterial indicator culture. ... This mixture was heated at 37oC for 5 minutes and after this time elapsed, 10 uL from the ADS turn was transferred to one of the sterile microfuge tubes held at 37oC, along with 3 drops of chloroform. ... Another 10 uL was taken from the ADS tube (after which the ADS tube was returned to the 37oC heat) and 3 one-hundred fold serial dilutions were made using the microfuge tubes sitting on ice (to make the IC tube).
Approximate Word count = 1672
Approximate Pages = 6.7
(250 words per page double spaced)
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