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Unadsorbed Phage Lab Report

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Objective
The purpose of this experiment was to determine the reversion rate and recombination rate of two mutant T4 bacteriophages, rII 29 and rII 31. Through recombination rate, the map unit between the two mutants was calculated.
Methods
Reversion rate of phage rII29 and phage rII 31
The reversion rate of either phage rII 29, or phage rII 31 was determined by the infection of permissive host E. Coli B, or with non-permissive host E. Coli K to either bacteriophages rII 29, or bacteriophage rII 31. This infection was plated using the soft agar technique. After incubation, the titer of revertants phage was determined by the number of plaques formed on E. coli K plates. The reversion rate equation was the following,
Frequency of reversion
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5 drops of chloroform were added to the mixture and incubated at room temperature for 15 minutes to kill, and lyse the bacteria cells which allows all the unadsorbed phages to be accounted for when determining the titer of unadsorbed phage with susceptible bacteria, E. Coli B. (Biology Dept.). 0.1 ml of E.coli B was added to the 10 fold dilution. Using soft agar technique, the unadsorbed phage were plated. After incubation, the titer of unadsorbed phage was determined by counting the number of plaques on the plate from the infection of single E. coli cells in the assay tube by a free phage particle before…show more content…
(Biology Dept.). 0.1 ml of E.coli K or 0.1ml of E.coli B was added to the 10 fold dilution. Using soft agar technique, the growth media mixture with E.coli was plated and incubated.
The titer of recombinant phage was determined with the number of plaques on the E.coli K plates and the titer of total phage progeny was determined with the number of plaque on the E.coli B plates.
Determination of wild type phage progeny
The titer of recombinant phage, where the mutant phage is reverted to wildtype, was done on E.coli K plates. The wild type phage progeny was determined by the plaques counted on the titer of E.coli K plates. The recombination rate equation was the
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