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homework10 - Viral Quantification Practice Homework...

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Viral Quantification Practice Homework Questions 2010 Against your advice (and the advice of probably every virologist on Earth), your roommate decides it’s not worth the effort to get a flu shot this year. Now he is laid up in bed for the next week with the flu, feeling approximately as though he has been run over by a train. You resist the urge to tell him ‘I told you so’, and instead decide that having this human virus factory lying on your couch presents the perfect opportunity to practice your skills as a budding virologist. You convince him to spit into a test tube, and then take the sample into lab for some analysis… 1. You measure the volume of the sample using a pipetman and determine it to be 0.5 mL, yet you have no idea what the viral titer of your sample is. How would you set up a plaque assay to determine the PFU of influenza virus in the sample? Take the 0.5 mL of your sample and do many 1:10 dilutions. Next, set up a monolayer of cells on a plate with agar. Take 0.1 mL of each dilution and transfer to their respective agar plate. Look for the plate with plagues that are easy to count (that look like holes) And then use the equation : PFU/ml = # plaques x 10 x dilution factor 2. You have just developed your plaque assay. You got the following results from infecting the cells with 1/10 volume (i.e. 100 ul from a 1 mL-containing tube) of each of your 10-
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