9.9 ml solvent 9.9 ml solvent 9.0 m solvent Tube 1 Tube 2 Tube 3 Dilution 100-fo

Question

9.9 ml solvent 9.9 ml solvent 9.0 m solvent Tube 1 Tube 2 Tube 3 Dilution 100-fold 10-fold Total dilution 100 x 100 x 10- 100,000 Figure 4-1. Preparing Serial Dilutions PROCEDURE 1. You should be practicing pipetting and preparing serial dilutions using water, before proceeding 2. A stock solution of the protein bovine serum albumin (BSA) has been prepared at a concentration of 20 milligrams per milliliter (mg/ml); you have a supply of this stock provided for you at your station. Bring a 30 ml supply of distlled water to your desk. Use the serial dilution technique to prepare 1.5 ml volumes of the various BSA concen- trations listed in Table 4-1. Write in the space provided how you will prepare each of the sample preparations. Be careful not to contaminate one concentration with solution from a different concentration. Save all dilutions, because each of these concentrations will be needed for Part C. V, 1.5 m

Explanation / Answer

Simple dilution is the part of cumulative dilution. In simple dilution, we dilute the sample in each step. Cumulative dilution is the multiplication of all the simple dilution.

As we can see in the figure that the tube 1 and 2 is diluted 100 times and tube 3 is diluted 10 times. All these dilutions are simple dilutions. When we calculate the overall dilution that the original sample has gone we call it cumulative dilution.

Calculation-

For the sample preparation, we will use following calculation

C1 is the concentration of the stock solution

C2 is the concentration of working solution

V1 is the volume of stock solution that we have used to make the C2 solution

V2 final volume.

In case of 20mg/ml we do not need to dilute the BSA as the stock solution is 20mg/ml

For 10mg/ml

C1 is the concentration of the stock solution =20mg/ml

C2 is the concentration of working solution 10mg/ml

V1 is the volume of stock solution that we have used to make the C2 solution we need to calculate

V2 final volume. 1.5ml

20*V1 = 10*1.5

V1 = 15/20=.75

So we need to add 0.75ml of 20mg/ml of solution in final volume 1.5ml to make 10mg/ml solution.

For 5mg/ml

here stock solution will be 10mg/ml

So 10*V1 = 5*1.5

V1 = 7.5/10

= 0.75ml

So we need to add 0.75ml of 10mg/ml of solution in final volume 1.5ml to make 5mg/ml solution.

For 2.5 mg/ml

here stock solution will be 10mg/ml

So 10*V1 = 5*1.5

V1 = 7.5/10

= 0.75ml

So we need to add 0.75ml of 10mg/ml of solution in final volume 1.5ml to make 5mg/ml solution.

For 2.5 mg/ml

here stock solution will be 10mg/ml

So 5*V1 = 2.5*1.5

V1 = 3.75/5

= 0.75ml

So we need to add 0.75ml of 5mg/ml of solution in final volume 1.5ml to make 2.5mg/ml solution.

For 1.25 mg/ml

here stock solution will be 10mg/ml

So 2.5*V1 = 1.25*1.5

V1 = 1.875/2.5

= 0.75ml

So we need to add 0.75ml of 2.5mg/ml of solution in final volume 1.5ml to make 1.25mg/ml solution.

I have done the calculation for you. Please fill the table in your own language.

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