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Electrophoresis (Online Course)

(based on 1869 customer ratings)

Author: Mary Ellen Koenn, MS, MT(ASCP), CLS(NCA)
Reviewer: Leslie Lovett, MS, MT(ASCP)

This course discusses the many types and applications of electrophoresis in the clinical laboratory. Commonly used terms are defined and procedures are described, including explanations of various electrophoretic patterns. A discussion on newer molecular diagnostic techniques that incorporate electrophoresis conclude this in depth look at electrophoresis.

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Continuing Education Credits

  • P.A.C.E.® Contact Hours: 1.5 hour(s)
  • Florida Board of Clinical Laboratory Science CE - General (Clinical Chemistry/UA/Toxicology): 1.5 hour(s)

Objectives

  • Explain the principle of electrophoresis and compare the factors that determine the mobility and rate of migration in electrophoresis.
  • Describe the role of the buffer in electrophoresis, types of support mediums, and the kinds of specimens used in electrophoresis.
  • Compare and contrast the following types of electrophoresis: routine electrophoresis, polyacrylamide gel electrophoresis, high resolution electrophoresis, capillary electrophoresis, isoelectric focusing, two-dimensional electrophoresis, pulsed field electrophoresis, and immunochemical electrophoresis.
  • Define each of the following terms: amphoteric, ampholyte(s), isoelectric point, sodium dodecyl sulfate, cathode, and anode.
  • Name and describe the stains used in electrophoresis.
  • Evaluate causes of irregular, distorted or atypical electrophoresis bands, along with correct sample application and proper care and use of buffers and support mediums.
  • Explain electroendosmosis and wick flow as encountered in electrophoresis and assess their effect on separated bands.
  • Discuss the theory behind and uses of densitometry.
  • Describe increased electrophoresis usage today and incorporation of electrophoresis techniques in molecular diagnostics.

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Course Outline

Click on a link in the outline to view a sample page from this course.

  • Electrophoresis
      • Introduction
    • Principle of Electrophoresis
      • Principle of Electrophoresis
    • Amphoteric
      • Amphoteric
    • Isoelectric Point
      • Isoelectric Point (pI)
      • Which statement is correct for a protein with an isolectric point (pI) of 7.0?
      • The pI of a protein is 9.2. This protein is placed in an electrical field where a buffer sets the pH at 10.0. Select the correct statement regarding t...
    • Mobility or Rate of Migration
      • Mobility or Rate of Migration
      • Rate of Migration
      • Which one of the following will slow down the migration of solutes in electrophoresis?
  • Electrophoresis and Buffers
    • Role of Buffers
      • Role of Buffers
    • Buffers and pH
      • Buffers and pH
      • Proteins in a buffer with the pH set at 8.6 will become anions and move to the positively charged electrode.
  • Specimens for Electrophoresis
      • Specimens
      • After reviewing the information on specimen samples for electrophoresis, select the one correct statement.
  • Electrophresis and Support Media
    • Types of Support Media
      • Types of Support Media
    • Agarose Gel
      • Agarose Gel
    • Polyacrylamide Gel
      • Polyacrylamide Gels
      • There are several different types of media that can be used in electrophoresis. Most methods today use a gel, cellulose acetate, agarose, or polyacryl...
      • Of the three types of gels discussed, agarose gels are stronger, thermostable, and transparent.
  • Electrophoresis Instrumentation
    • Equipment
      • Electrophoresis Equipment
    • Automation
      • Automation
      • Automated electrophoresis systems only include automated reagent addition, electrophoresis of sample, staining of separated fragments, and detection o...
  • Types of Electrophoresis
      • Types of Electrophoresis
    • Routine Electrophoresis
      • Routine Electrophoresis
    • High Resolution Electrophoresis (HRE)
      • High Resolution Electrophoresis (HRE)
    • Polyacrylamide Electrophoresis (PAGE)
      • Polyacrylamide Electrophoresis (PAGE)
      • Denaturing Polyacrylamide Gels
    • Isoelectric Focusing (IEF)
      • Isoelectric Focusing (IEF)
      • IEF Advantages and Applications
    • Capillary Electrophoresis
      • Capillary Electrophoresis (CE)
      • CE Advantages and Applications
    • Two-Dimensional Electrophoresis
      • Two-Dimensional Electrophoresis
      • Two-Dimensional Electrophoresis Advantages and Applications
    • Immunochemical Electrophoresis
      • Immunochemical Electrophoresis
      • Immunofixation Electrophoresis
      • Electroimmunoassay Electrophoresis
    • Pulsed Field Electrophoresis
      • Pulsed Field Electrophoresis
      • In isoelectric focusing, the basis of separation of solutes is different than the other types of electrophoresis. Which statement below correctly desc...
      • Currently there has been a revitalization in the clinical usage of electrophoresis. Previously, methods were primarily used to separate proteins in bl...
      • Sodium dodecyl sulfate is added to polyacrylamide gels to denature the proteins in the sample and enhance their separation.
  • Visualization and Detection
    • Visualization and Detection Methods
      • Visualization and Detection Methods
      • Stains and Dyes
      • Densitometry
      • Labeled Probes
      • Which statements below are correct descriptions of visualization and detection methods used in electrophoresis?
  • Technical Considerations and Electrophoresis Troubleshooting
    • Topics for Consideration
      • Technical Considerations and Electrophoresis Troubleshooting Topics
      • Sample Application
      • Buffers
      • Support Media
      • Electroendosmosis
      • Wick Flow
      • Troubleshooting Irregular, Distorted or Atypical Bands
      • Which statements below are associated with electroendosmosis?
      • Wick flow is caused by movement of the buffer ions into the medium when there has been a loss of moisture in the medium due to heat generation.
  • Resurgence of Electrophoresis
    • Resurgence of Electrophoresis
      • Resurgence of Electrophoresis
      • Electrophoresis and Molecular Diagnostics
      • Blotting Techniques
      • Uses of CE in Molecular Diagnostics
  • References
      • References

Additional Information

Level of instruction: Intermediate

Intended Audience: Clinical laboratory technologists and technicians, and other health care personnel who have an interest in this subject matter. This course is also appropriate for clinical laboratory science students and pathology residents.
 
Author information: Mary Ellen Koenn, MS, MT(ASCP), CLS(NCA) is an associate professor at West Virginia University School of Medicine in the Medical Technology Program. During her career as a medical technologist and educator, she has been a laboratory supervisor and manager and has held several teaching positions. She is active in the American Society for Clinical Laboratory Science (ASCLS), resulting in a nomination as ASCLS Member of the Year in 2006. She is also a member of the American Association for Clinical Chemistry. She is the author of numerous articles for laboratory publications and textbook chapters and is a frequent presenter at laboratory seminars and workshops. Ms. Koenn holds a Master of Science degree in Medical Technology.
 
Reviewer information: Leslie Lovett, MS, MT(ASCP) is the Clinical Education Coordinator of the Medical Laboratory Technology Program and a professor at Pierpont Community and Technical College in West Virginia.  She holds a MS degree in Medical Technology with classes toward a PhD in Pharmacology and Toxicology.

Course Description: This course discusses the many types and applications of electrophoresis in the clinical laboratory. Commonly used terms are defined and procedures are described, including explanations of various electrophoretic patterns. A discussion on newer molecular diagnostic techniques that incorporate electrophoresis conclude this in depth look at electrophoresis.





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