G418 disulfate (FW 692.7) is an antibiotic used to select transfected mammalian cells in tissue culture. The standard working concentration for selection is 400 µg/mL of active ingredient in culture medium. The bottle states that it contains 71% active ingredient. You need to make a 100x stock in medium and adjust the pH before you apply it to cells. How much powder should be added to make 10 mL of stock solution? How much do you add to a 2 mL culture dish?
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- How 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.A vial of Doxorubicin reads 0•5g per vial. Instructions say to reconstitute each 12mg with 2•5ml of NS. How many ml of NS will be needed to reconstitute the vial of the recommended concentration? please show workingYou have an order for 1 gram of Cefazolin in D5W 100 ml. You have added 5 ml of sterile water to the 1 gram vial to reconstitute powder. However the recommended manufacturer’s diluent amount is 10 ml of sterile water for a final concentration of 100 mg/ml. How would reconstituting the vial with 5 mls affect the concentration and the final calculated dose?
- A culture of S. cerevisea has an overnight OD of 4.5 (1.0 OD is approx 1.0x107 cells/ml) You will be plating 100µl onto agar and want the final count of colonies on the plate to be around 300 colonies. How much of the 4.5 OD culture must you use to get a 500µl subdilution (with sterile water), so that you have diluted enough to get approx 300 colonies per 100ulYou 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.Nutrient Agar (NA) is a general purpose medium used for the cultivation of a wide variety of non- fastidious microorganisms (Merck, 2000). Its ingredients are listed in Table 2. You are tasked to prepare 300ml of NA, determine the amount of ingredients and write in column 3 in Table 2. Show your calculations. *A sample calculation was made for you. Peptone = 300 ml x 0.5% = 1.5 g Table 2. Medium Composition of Nutrient Agar (NA) with the recommended proportions (Merck, 2019) Ingredients In % In Grams/300ml Peptone 0.5 *1.5 Meat extract 0.3 Agar 1.5 Distilled Water As needed
- You have an order for 1 gram of Cefazolin in D5W 100 ml. You have added 5 ml of sterile water to the 1 gram vial to reconstitute powder. However the recommended manufacturer’s diluent amount is 10 ml of sterile water for a final concentration of 100 mg/ml. How would reconstituting the vial with 5 mls affect the concentration and the final calculated dose. Please answer with explanation ASAP. I will really upvote. ThanksEach liter of an intranasal antibiotic solution contains 160 mg of gentamicin. You have 2-mL gentamicin vials that contain 40 mg. How many vials will you need if you are to prepare three liters of the antibiotic solution?What is the amount (mL) of pre-culture (108 cell/mL) necessary to inoculate (to add) in a 2-liter culture media for a concentration of 6.5 x 106 cells/mL?
- A bacterial suspension is standardized using a 0.5 McFarland Standard. 1 mL of that bacterialsuspension is aseptically transferred to a 250-mL volumetric flask and then diluted to volume withsterile water. After this is mixed, a 2.0-mL portion of the resulting suspension is taking andtransferred aseptically to a 50-mL volumetric flask, where it is diluted to volume with water. Thebacterial suspension is mixed and then prepared for use in product testing.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?There are many ways you can accomplish a 1000-fold dilution. Propose 3 different serial dilution methods that will accomplish a dilution with a 1:1000 dilution factor.