Understanding the Basics of Electrophoresis
Master the process that separates charged particles in a fluid using a field of electrical charge in this free course.Publisher: NPTEL
CertificationView course modules
In this free online course, you are introduced to the world of electrophoresis. Discover whether this technique is suitable for the separation and analysis of complex biological samples. Discuss the four different stains that are available for the detection of proteins in polyacrylamide gels. You will be introduced to the two types of gels that are cast in a vertical gel electrophoresis and see how the vertical system performs differently to a horizontal gel system. You will discover if a sypro tangerine stain can be used to detect blotted protein bands. Demonstrations, problems, examples and assessment questions in this instructor-led video-based course, are designed to provide you with a comprehensive foundation in electrophoresis.
The material introduces you to the definition of horizontal and vertical gel electrophoresis and the different types of stains that are used to detect proteins in polyacrylamide gels. Then, the electrophoresis in the horizontal gel system, which is performed continuous with both electrodes and gel cassettes submersed within the buffer, will be discussed. You will explore the fact that ammonium persulfate reagent is used as an initiator in the acrylamide polymerization process, whilst sodium dodecyl sulphate is a reagent that is used to denature and provide a negative charge to the protein. You will discover the fact that the agarose reagent is used in the preparation of the horizontal gel for DNA analysis and the additional accessories that are needed for casting the agarose gel. It will be clarified that there are two platinum electrodes in the electrophoresis chamber that are part of the horizontal gel electrophoresis system.
Next, you will learn that the complex biological samples are efficiently resolved in 2-D gel electrophoresis, which involves the combination of charge and molecular weight to provide much greater separation in comparison to the use of the individual property. You will gain insight into the fact that when conducting the agarose gel electrophoresis experiment on the native page, the sample that is prepared in the loading dye does not contain detergent or denaturing agent and as a result, the sample runs based on charge. You will then discover that in the horizontal polyacrylamide gel electrophoresis, the complex biological sample is resolved according to its charge and then moved to the counter charge electrode. You will be taught that in the urea page method, insoluble protein is dissolved in urea and samples are separated based on their subunit mass. Finally, you will study reagents in gel electrophoresis, staining and image analysis, variants of gel electrophoresis, and the components of gel electrophoresis. This course will be of interest to biotechnology students or those professionals in experimental biotechnology wanting to hone their technical knowledge.Start Course Now
Upon successful completion of this course, you should be able to:
- Discuss the components of gel electrophoresis
- Explain the steps in the staining procedure
- List the typical apparatus in an agarose gel electrophoresis experiment
- Describe the different variants of gel electrophoresis
- Discuss supporting media that is used in zone electrophoresis
- Explain the steps in the 2-D gel electrophoresis process
- Describe how the agarose gel electrophoresis is used to dissolve the DNA
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