Real-Time PCR (Online Course)

(based on 503 customer ratings)

Author: Ryanne Wolfe, MS, MBA
Reviewer: Dana L. Van Laeys, MA, CLSpMB, MT, CLS

This course provides a brief overview of DNA and RNA structure and the principles of polymerase chain reaction. It then proceeds to discuss the principles and procedures for real-time PCR, highlighting the advantages over traditional PCR methods and the applications in the clinical setting.

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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 (Molecular Pathology): 1.5 hour(s)

Objectives

  • Compare and contrast the structure and functional regulation of DNA and RNA.
  • Discuss the principles of the polymerase chain reaction (PCR) process.
  • Define the detection techniques for the measurement of nucleic acids by real-time PCR.
  • Understand the evolution of traditional PCR to real-time PCR, and the advantages and limitations of both methods.
  • Discuss the applications of real-time PCR.

Customer Ratings

(based on 503 customer ratings)

Course Outline

  • Historical landmarks in Molecular Diagnostics
      • Historical Landmarks in Molecular Diagnostics
  • Composition and Formation of DNA and RNA
      • Molecular Composition of DNA
      • Molecular Composition of RNA
      • Formation of DNA and RNA Strands
      • Which two of the following characteristics are common to both RNA and DNA?
      • The DNA strand ending with a phosphate group is the 5' end.
  • Extraction and Isolation of Genetic Material
      • DNA Isolation
      • Automated Vs. Manual Extraction
      • If a cocktail that includes detergents, enzymes, and salts is used for DNA isolation, which one of these ingredients will pull the DNA out from the sa...
  • Principles of Polymerase Chain Reaction (PCR)
      • PCR Introduction
      • Reaction Mixture
      • DNA Primers
      • DNA Polymerase
      • What temperature is considered optimal for Taq polymerase activity?
    • Steps of PCR
      • Denaturaton
      • Annealing (Hybridization)
      • Extension
      • Repetitive Cycling
      • Diagram of PCR Process
      • What is the first step of the PCR reaction?
    • PCR Results
      • Analyzing PCR Results
  • Reverse Transcriptase PCR (RT-PCR)
      • Reverse Transcriptase PCR (RT-PCR)
  • Real-Time PCR
      • Introduction to Real-Time PCR
      • Advantages of Real-Time PCR
      • Stages of Real-Time PCR
      • Diagram of the Real-Time PCR Stages
      • Detection Methods
      • FRET
      • In real-time PCR procedures, fluorescence resonance energy transfer (FRET) is the process that allows for probes to emit fluorescence only after hybri...
      • When is data collected in a real-time PCR procedure?
    • Real-Time RT-PCR
      • Real-Time Reverse-Transcriptase- PCR
    • Real-Time PCR Results
      • Quantification
      • Real-Time PCR Computer Results
      • During which stage in real-time PCR can the fluorescent signal start to be recorded?
  • Applications of Real-Time PCR
      • Laboratory Applications
      • Multiplexing
      • Microbe Detection
      • SNP Detection
      • Viral Load
      • The best way to evaluate single nucleotide polymorphism (SNP) is by melting curve analysis
      • Which is not an application of real-time PCR?
  • Conclusion
      • Conclusion
  • References
      • References

Additional Information

Level of instruction: Intermediate

Intended audience: Medical laboratory scientists, medical technologists, and technicians, This course is also appropriate for clinical laboratory science students and pathology residents. 
 
Author information: Ryanne Wolfe holds an MS in biotechnology and an MBA from the University of Maryland. Ryanne has worked in the health care field for eight years, most recently in the laboratory at St. Christopher’s Hospital for Children in Philadelphia, Pennsylvania.
 
Reviewer information: Dana L. Van Laeys, MA, CLSpMB, MT, CLS is the Education Coordinator for Molecular Diagnostics and Immunology in the Clinical Laboratory Science Program at Saint Luke’s Hospital in Kansas City, Missouri, a position that she has held since 1998. She has 17 years of experience in molecular diagnostics and flow cytometry. She has been a presenter at ASM, CLEC, and AABB conferences. Ms. Van Laeys holds a Masters degree in Adult Education and Distance Learning. 
 
Course information: This course provides a brief overview of DNA and RNA structure and the principles of polymerase chain reaction. It then proceeds to discuss the principles and procedures for real-time PCR, highlighting the advantages over traditional PCR methods and the applications in the clinical setting.




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Real-time PCR Stages