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Repeat Steps for Buffer B see previous experiment or any other buffer.

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Tabulate the data and analyse the result. Solute is a substance which is present in small quantity in a solution. Solvent is a substance which is present in large quantity in a solution. Many solutions used in biochemistry are prepared by the dilution of a more concentrated stock solution.

In preparing to make a dilution or series of dilutions , you need to consider the goal of the procedure. This means that you need to consider both the desired final concentration and required volume of the diluted material.

A simple equation allows the dilution to be calculated readily: illustration not visible in this excerpt Where, C1is the concentration of the initial solution; V1is the volume of the initial solution available to be used for dilution this may not be the total volume of the initial solution, and instead may be a small fraction of the initial solution , C2is the desired final concentration, and V2is the desired final volume. In most cases, the initial concentration and the final concentration are either known or are chosen in order to work correctly in the experiment being planned.

The final volume is usually an amount that is chosen based on the amount required for a given experiment. This means that at least three of the required terms are either known or can be chosen by the experimenter.

This leaves V1 as the unknown value i. This is a problem: 0. You have two choices: change the final volume i. In many cases, while the final concentration is important, the final volume is not as in the previous paragraph.

In these cases, do what was explained in this example: use a convenient dilution: a dilution that involves volumes that are easily pipetted. Pipetting 1. In some cases, you may not know the actual starting concentration.

If, for example, you need to measure the enzyme activity in a sample, and you find that the activity is too high to measure accurately, you will need to dilute the starting material.

As long as you keep track of the concentration ratio in all of your dilutions, you can easily determine the enzyme activity in the initial solution, even though you cannot measure it directly. Concentration ratios are frequently of considerable value.

In each case the concentration of the final buffer is one-tenth that of the original. Simply performing a dilution of the stock solution then gives the appropriate final concentration of each component.

The stock solution of buffer is typically called a 10x stock, because it is ten-times more concentrated than the final, useful buffer. The dilution mentioned is performed by taking one part of the initial solution, and adding nine parts of solvent usually water.

This results in a final concentration that is ten-fold lower than the original. It is usually done if the chemical concentration is too high than the desired composition.

Thus, serial dilution simply means a series of repeated dilution performed on the same chemical basically to change its concentration. After performing the dilution, we need to know how much difference are the diluted chemical and the initial, undiluted ones. There are several benefits of performing serial dilution. Serial dilution comes in handy when the solution is too concentrated to be used in experiments or ingredients preparation.

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This is to ensure that the exact concentration can be obtained for the experiment to become success. Other than that, the diluted solution from this serial dilution can be used to count the concentration of the actual solution. By knowing the dilution factor of certain solution, the calculations of the concentration become easier and systematic. This method is applicable in several fields, not only in chemistry. Serial dilution is a simple yet efficient technique to determine the number of cells or organisms in a concentrated sample.

First, take a portion of the sample and does serial dilution on it. Repeat the steps until the cells can be observed under the microscope when the diluted sample was observed.

Then, count the dilution factor and times it with the actual volume of the sample. Other than that, medicine administration require suitable dose for each patient with variable needs.

This is where serial dilution is useful. The final volume is equal to the aliquot volume plus the diluent volume: 0. Additional examples on dilution and serial dilutions A. One mL of a sample was mixed with 99 mL of buffer. One mL of this was plated using the pour plate method in nutrient agar.

After incubation, colonies were present on the plate. How many colony-forming units were present per mL of the original sample? The dilution used was [Abbildung in dieser Leseprobe nicht enthalten]. One mL of the dilution contained colony-forming units. How much did one mL of the original sample contain? Obviously more than colony-forming units! Multiplying by the dilution factor: colony-forming units[Abbildung in dieser Leseprobe nicht enthalten] of original sample.

Either way, the answer you get is the same. One mL of a sample was mixed with 99 mL buffer. One-tenth of amL of this was plated on nutrient agar. In this case, the mathematics of the dilutions is the same, but the number of colonies counted on the plate the number of colony-forming units in only 0. Free Preview. Includes step-by-step, easy to follow protocols Provides key tips and expert implementation advice to ensure quality results Provides up-to-date techniques along with a brief theoretical introduction for each section see more benefits.

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About this book This book is a practical guidebook in biochemistry, for medical as well as life sciences' students. Show all. Pages Blood Collection and Preservation Kumar, Vijay et al.

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Quality Control in Laboratory Kumar, Vijay et al. Fundamentals, Methods and Clinical Applications. Examkrackers Mcat Biology: Jonathan Orsay. Textbook of Biochemistry with Clinical Correlations.

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