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Assignment 1 for MCB 3020L Spring 2014 (due 2/24 or 2/26) Instructions: * Your answers have to be typed and double spaced. * Put the date, your name and your TA’s name on the cover page of this assignment. * “Give a detailed explanation” means don’t just give a one sentence answer. * Don’t forget to include your calculations and references!  Late assignments are not accepted for grading.

Questions: 1) Assume that you measure the OD of two E.coli cultures: culture A and culture B. The spectrophotometer gives you a reading of 2.2 for culture A and 0.02 for culture B. What would you do to determine the concentration of culture A and B and why? Give a detailed explanation!

For culture A the OD reading is 2.2. This must never exceed 1, so a dilution is required.
For culture B the OD reading is 0.2. You would calculate the reading by the factor 1x10^8 cells/mol of E.Coli

2) The OD of an E.coli culture is 0.46. You transfer 60µl of this culture into 5.94ml of medium. You continue this same dilution one more time. You transfer 25 µl of your last dilution onto an LA plate. After 24 hours of incubation, you observe 12 colonies on this last plate. A) Did you expect this result? If not, name at least three reasons for this result giving a detailed explanation for each reason. B) What would you do to prove your theory and why? A)

3) A friend of yours asks you to help him/her draw two bacterial growth curves. He/she gives you the following information: the TA started both cultures (one was labeled culture “X” and the other culture “Y”) at 8:30am using the same overnight culture to inoculate fresh medium. After inoculation of the medium the OD was 0.14 for both cultures and both cultures were incubated at 37 oC. During class the following OD

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