1 ml A 1 ml B 1 ml ? Original 0000 1 2 3 4 5 9 ml 9.9 ml C 9.9 ml D 1 ml Plated on each 1 2 3 4 5 CFU TNTC 8963 852 98 9 1. Fill in the volume transferred for A and B. A = _____ B= _____ 2. Fill in the volume of diluent that is present in C and D. C= _____ D= _____ 3. Calculate the serial dilution factor for tubes 1-5. 1= _____ 2= _____ 3= _____ 4= _____ 5 = _____ 4. Calculate the plate dilution factor for plates 1-5. 1= _____ 2= _____ 3= _____ 4= _____ 5 = _____ 5. Choose the plate(s) that have the CFU within the statistically reliable range. 6. Using the CFU from your answer for question #5-calculate the OCD.
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Please note that for the dilution problems, use only the following diluent volumes (0.9 mL, 9 mL, or 9.9 mL). Answers should be in terms of exponents. 1. Draw TWO ways of making a 10^-6 dilution. Indicate all transfer volumes and diluent volumes. Include IDF & TDF values. 2. One mL of a sample that had been diluted to 10^-8 was spread on a TSA plate. The plate yielded 80 CFU. What is the original cell density (OCD)? Show calculation & provide appropriate units. 3. 0.1 mL of a sample that had been diluted to 10^-7 was spread on a TSA plate. The plate yielded 150 CFU. What is the original cell density (OCD)? Show calculation & provide appropriate units.
Farhan A.
A. The standard plate count method is performed with dilutions of the original sample. 0.1 ml of dilution was plated on each petri dish and pour plates were made. The 1/10^3 dilution had 250 colonies. The 1/10^4 dilution had 25 colonies. What was the CFU/ml of the original sample? B. You take 1 ml of an original sample, dilute it with 9 ml of buffer and mix them together. You then take 1 ml of the diluted sample and mix it with 99 ml of buffer. You take 1 ml of the last diluted sample and mix it with 99 ml of buffer. How much is your sample diluted at the end?
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Use the following numbers to complete the exercise on serial dilutions. You have counted three plates that have colonies growth on them. Note the dilution factors and calculate the approximate number of bacteria in the original volume of the test tube. -6 -7 -8 644 111 27 1702 289 13 357 85 9 OCD = CFU/D X V OCD = original cell density in scientific notation/ml CFU = colony forming units D = dilution written on the tube V = volume transferred onto the petri dish
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