5 Prelab Reading Dilutions and Standard Curve

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Michigan State University *

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65.105

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Chemistry

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Feb 20, 2024

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docx

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Pre-Lab Reading: Dilutions Objectives: This reading should help you, by the end of this experiment, to be able to: 1. Correctly measure fluids using a pipette 2. Carry out a serial dilution 3. Use data from a spectrophotometer to create a standard curve showing algal concentration 4. Identify morally relevant facts in ethics issues in conducting research, critically evaluate those facts in order to develop a resolution The purpose of this laboratory exercise relates directly to the experiments you will be conducting throughout the semester. In today’s lab, you will learn how to determine the concentration of algae in a solution of saltwater. This is critical for working with plankton, as the algae are the food source that they need in order to survive. If your algae are not concentrated enough, the plankton will starve and will die in large numbers. If your algae are too concentrated, this can affect the swimming and breathing ability of the plankton. Therefore, it is important to know just how many algal cells are in the solution that you are using to feed the plankton. In order to know how much to feed your copepods each week, you will need to use a standard curve to determine the concentration of the algae. Today we are going to practice making a standard curve and learning how to use one to determine concentrations. We will do this in five steps: 1. Make a serial dilution 2. Use a spectrophotometer to find the absorbance of our solutions 3. Enter data points in Excel 4. Graph a standard curve 5. Find the concentration of an unknown using the standard curve Dilutions Orange juice is a great example of dilution – when you make orange juice from a can, the stuff in the can is called “concentrate”. You then dilute it with water. Serial dilutions are made with the same general idea, but a slightly different process. To make a serial dilution, you begin with a stock solution. A stock solution is the concentrated solution which is being diluted. Next, you need to know your dilution factor , which is a number that describes the strength of the dilution . For example, a dilution factor of 10 means a 1:10 dilution of the stock solution. To
1 ml of stock solution 9 ml of water calculate the dilution factor, you take the volume of the stock solution that you are going to use and divide this by the total volume of the entire solution. volume of stock solution youare using total volume your dilution For example, if you are doing a 1:10 dilution, you would take 1ml of your stock solution and add 9ml of water for a total volume of 10ml. 1:10 dilution In many cases, a dilution factor of 10 or 2 is used to dilute a stock solution and create a series of less concentrated solutions. This process is referred to as making serial dilutions. A serial dilution is a sequential set of dilutions, where each solution acts as the stock for the next solution. Essentially, it is just the process of making dilutions with the same dilution factor over and over. To carry out a 1:10 serial dilution, assume that you start with 11 ml of your stock solution. To make a 1:10 dilution, take 1ml of your stock and add 9ml of water. This will give you a new stock solution that has been diluted by a dilution factor of 10. You now take 1ml of this new stock solution and add 9ml of water. This gives you yet another new stock solution that has been diluted by a dilution factor of 10. This process is repeated until you reach the total dilution you were looking for. The image below may help understand the process of serial dilutions.
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