3. 4. Draw a picture of a ten-fold serial dilution. Please use 10ul ( 9ml of solution. Show a picture of what a serial dilution would thousand-fold. You can draw this and take a picture.
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- You have a sample at 50 ng/ul and you would like to load 400ng of this sample on a lane of an agarose gel. You also have TE buffer as diluent and 6x loading dye. Your total sample should be 12ul. Calculate the amounts of each reagent necessary to prepare this sample for gel loading.Prepare the following LB media with antibiotic added. A 100ml of LB media with 25 micro/ml of Amp and 100micro/ml of Kan final concentration. A 100ml of LB is provided and amp of Kan stock at 100 mg/ml and 50 mg/ml are also provided. Find how much each antibiotic stock solution is needed to add 100ml of LB to reach desired antibiotic concentration.1) You have been asked to make up four 1.5% agarose gels at 30 ml each. Ethidium Bromide is to be added at 0.5 ug/ml and you have a bottle at 10ug/ml. Write down step-by-step protocol of how to proceed. 2. How would you make up one liter of 1X TAE buffer using a 25X stock?
- Explain how gel filtration chromatography works. What type of gel will you used when the protein size is 2500 Da? Explain.100ml of LB media with 25 μg/ml of Amp and 100 μg/ml of Kan final concentration. You have 100ml of LB provided and Amp and Kan stocks at 100 mg/ml and 50 mg/ml provided. Determine how much of each antibiotic stock solution you need to add to 100ml of LB to reach desired antibiotic concentration.The SDS-PAGE protocol requires 0.5L of running buffer for the gel apparatus. The stock running buffer comes as a 20x concentrate. How should you prepare your running buffer? 50ml 20x stock + 450ml deionized water 450ml 20x stock + 50ml deionized water O 250ml 20x stock + 250ml deionized water 25ml 20x stock + 500ml deionized water 25ml 20x stock + 475ml deionized water 2ml 20x stock + 500ml deionized water O 100ml 20x stock + 400ml deionized water
- 1. (2.5 pts) A pure bacterial culture of unknown concentration was diluted to determine the concentration of viable bacteria in the original culture. Serial dilutions were performed as diagrammed below. Each dilution tube contained 400 ul of diluent and 100 ul was transferred into each tube. TSA plates were inoculated with 100 µul from the last three dilution tubes. a. What is the dilution between each tube shown in the diagram below? Express your answer as a ratio. b. What is the total dilution of tube number 5? Express your answer as a ratio. c. What is the concentration of viable bacteria in the original culture? Express your answer using scientific notation and the units CFU/ml. d. What is the concentration of viable bacteria in tube number 2? Express your answer using the units CFU/ml. e. If you inoculated a TSA plate with 250 µl from tube number 5, how many colonies would you expect to see after the plate was incubated? 1 2 3 5 423 80 13 Number of colonies1. Why is necessary to remove fat and tendons from the heart sample? 2. Why is necessary to completely grind the beef? 3. Why is necessary to balance the homogenate tubes for centrifugation? 4. How do you prepare a 100mL of 0.1 M phosphate buffer? 5. From the anterior buffer, how do you make 100mL of 0.05 M? 6. Calculate the amount of ammonium sulfate necessary to get a 20% solution? 7. Which is the importance of dialysis?2.1The number of bacteria in the samples need to be quantified. The laboratory uses a specificprotocol where the dilution factor on the spread plates must be:10-6; 10-7; 10-8Describe how you would prepare the required dilution series using the least amount of dilutionwater. 2.2 Calculate the amount of bacteria in the sample from the following results (show the steps in the calculation): Dilution factor Count10-6 27110-7 3510-8 21
- Serial dilution = previous dilution x dilution 11 mL [2] 3 mL Colony count: 100 0.1 mL sample [1] 9.9 mL 5 mL 0.1 ml. Cokony count Dilution: 10-1 10-3 [3] 150 Dilution factor (DF): [4] [5] [6] Compute for the CFU/mL of the sample. Show your solution and express your final answer to 2 significant figures.3) With our 12.5 ug/mL solution, we will make serial dilutions with a dilution factor of 2. We want to make 1 mL of 1/2, 1/4, 1/8, 1/16, 1/32, and 1/64 dilutions. Draw a diagram describing how you will conduct the serial dilutions and insert it below. The 1/2 dilution (tube 2) will be made from the 12.5 ug/mL solution (tube 1). The 1/4 dilution (tube 3) will be made from the 1/2 dilution (tube 2LHow would you make two-fold serial dilutions such that the last tube is a 1:32 dilution of the original, concentrated material? Assume that you need to have at least 500 µl of each dilution, and you should be able to perform the dilutions in microfuge tubes with a maximum capacity of 1.5 ml.