Real-Time PCR (Online Course)

(based on 417 customer ratings)

Coauthors: Heather MacDonald, M(ASCP), MB(ASCP) and Ryanne Wolfe, MS, MBA
Reviewer: Alex Casapu, MBA, MLS(ASCP)CM

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 417 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: Heather MacDonald, M(ASCP), MB(ASCP) has over 10 years of clinical laboratory experience and oversees a Molecular Diagnostics Laboratory. She is currently the Advanced Diagnostics Manager for Children's Healthcare of Atlanta in Georgia. Along with performing routine diagnostic assays, implementing laboratory-developed qualitative and quantitative molecular assays (both singleplex and multiplex) is her primary focus. Heather has published numerous articles and presents her research at national and international meetings. She has also worked with numerous corporations to bring commercial assays to market.
Coauthor 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: Alex Casapu, MBA, MLS(ASCP)CM, has 20 years of experience as a medical laboratory scientist, section supervisor, and laboratory manager. He is the former Director of Clinical Laboratory Technology Program at Georgia Piedmont Technical College. He is currently a Program Director at MediaLab, Inc. Alex holds BS degrees in Biology and Medical Technology from Clark Atlanta University and a MBA from the University of Georgia.
 
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