Genetics - MIT Department of Biology 7.02 Experimental Biology Communication Spring 2005 7.02/10.702 Spring 2005 7.02/10.702 Microbial Genetics

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MIT Department of Biology 7.02/10.702 Spring 2005 7.02/10.702 Microbial Genetics Exam Study Questions These questions—adapted from old exam questions--are meant to help you prepare for the 7.02/10.702 Genetics exam on March 8 th , 2005. The exam will likely contain 4 questions based on both lecture and laboratory material. The exam is CLOSED BOOK and CLOSED NOTES , and w ill be held in the lecture hall, during normal lecture time. These study questions are not meant to be exhaustive, but should give you an idea of what topics you should study. We strongly urge you to work through these questions before looking at the answers, and bring any questions to your Undergrad TA, Grad TA or one of the Instructors. Answers to these questions will be available on the 7.02/10.702 web site! 1
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7.02/10.702 Spring 2005 Question 1 You are handed an undiluted culture of pNK/KBS1 E. coli , and are told that it contains 4 x 10 11 cells/L. You also know that 1 OD550 of pNK/KBS1 = 1 x 10 8 cfu/mL. a) You want to take an OD550 of this culture. By what factor do you need to dilute the cells to ensure an accurate spectrophotometer reading of 0.25? SHOW YOUR CALCULATIONS. b) Complete the following sentence to describe how you would make 1 mL of diluted culture with an OD550 of 0.25: I would add __________ microliters (µL) of culture to __________ microliters (µL) of dilutant to obtain a final volume of 1 mL. c) If you made a 1:10,000 dilution of a culture with an OD550 of 0.25, and plated 100 µL of that diluted culture onto an LB plate, how many colonies would grow on the plate? SHOW YOUR CALCULATIONS. To set up an experiment, you mix 3 ml of undiluted pNK/KBS1 cells (titer of 4 x 10 11 cells/L) with 2 ml of P1 phage with a titer of 10 8 pfu/mL. d) Determine the MOI of this experiment. SHOW YOUR CALCULATIONS. e) Circle the experiment that the MOI calculated in part d) is more appropriate for: making P1 transducing lysates OR P1 transduction Explain your answer in two to three sentences by stating why that experiment requires that type of MOI (i.e. what do you want to happen/not happen in the experiment, and how does this kind of MOI ensure that?). 2
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7.02/10.702 Spring 2005 Question 2 During 7.02 lab, Andrew and Kate isolated an Ara- mutant, and observed dark blue colonies when they patched this mutant on both LB X-gal Kan plates and LB Ara X-gal Kan plates. From this data, they concluded that they had succeeded in creating an ara::lacZ translational fusion, mostly likely to the araC gene. a) The symbols shown below represent the protein product of the lacZ gene and the protein product of the wild type araC gene ( araC+ ): lacZ encoded araC +-encoded protein protein Draw a diagram of the protein product of an araC::lacZ translational fusion, and label the parts of your diagram with the appropriate protein names. b) Explain
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This note was uploaded on 06/11/2008 for the course BI 341 taught by Professor Driesen during the Spring '08 term at Portland State.

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Genetics - MIT Department of Biology 7.02 Experimental Biology Communication Spring 2005 7.02/10.702 Spring 2005 7.02/10.702 Microbial Genetics

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