One Step Growth Curve Of T4 Bacteriophage In E.coli

Many techniques can be employed in order to understand how viruses enter,
replicate and assemble in cells. In 1939, Ellis and Delbrück developed a
method to study a single round of virus multiplication using a one step growth
curve. This method is possible due to the synchronization of replication of
phage in the sample (Flint, 2002).

A one step growth curve experiment begins when bacteriophage are infected
with a very large number of phage particles. The large number of phage
particles ensures all bacterium are rapidly infected. This high level of infection
is called multiplicity of infection (MOI) and can be achieved with a phage to
host ratio of 5 to 10 plaque-forming units (PFU) per cell (Flint, 2000). The
phage/cell mixture is then diluted synchronizing the infection. As the infection
of bacteriophage is synchronized, the interaction of virus with the cell
population can be seen as a single interaction between phage and cell. In
order to visualize the infection over time, samples of the phage/cell mixture are
removed at intervals, plated and then quantified using the plaque assay
(Madigan, 2000)

It is possible to divide the multiplication cycle of viruses into five periods. The
first period is the eclipse p

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.

Sample Calculation for bacteria/mL ADS

Using the UP, FC and illustrative IC values the burst size was determined
to be 3 phage particles per bacterium.
 Some topics in this essay:
Ellis Delbrück, ADS Bacterium, Using FC, Results PFU, Calculation PFU/mL, Dilution Factor,
burst size, Unadsorbed Phage, , growth curve, latent period, ADS PFU, phage particles, step
growth curve, ads tube, step growth, t4 bacteriophage, sample calculation, flint 2000, ic plates,
Sample Calculation, calculation burst size, sterile broth diluent, using sterile broth, bacteria vs
elapsed,
 
   
   
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