Real-Time polymerase chain reaction( PCR)

Real-Time polymerase chain reaction( PCR) is organized by MediaLab, Inc.

Course Overview:
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.

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.

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
    • Nucleic Acid Isolation
    • Automated Vs. Manual Extraction
    • There are several methods for extracting nucleic acids (DNA and/or RNA}) from samples. The most common protocols are manual or automated which essenti...
• Introduction to Polymerase Chain Reaction (PCR)
    • PCR Introduction
    • Steps of PCR - Denaturation
    • Steps of PCR - Annealing (Hybridization)
    • Steps of PCR - Extension
    • Repetitive Cycling
    • Overview of PCR Basic Principles
    • What is the first step of the PCR reaction?
• Real-time PCR
    • Conventional PCR vs. Real-time PCR.
    • Real-time PCR Components
  • Primers
    • Primers
  • Polymerase
    • Polymerase
  • Enzyme Cofactors, dNTPs, and Salts/Buffers
    • Enzyme Cofactors, dNTPs, and Salts/Buffers
    • Reverse Transcriptase PCR (RT-PCR)
  • Thermocyclers and Detection Methods
    • Thermocyclers
    • Detection Methods
    • Phases of Real-Time PCR
    • During which phase 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
    • One of the best ways to evaluate single nucleotide polymorphism (SNP) is by melting curve analysis
• Conclusion
    • Conclusion
• References
    • References