Course Outline
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- 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
- Primers
- Polymerase
- Enzyme Cofactors, dNTPs, and Salts/Buffers
- 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
- Fill in the blank: The PCR type used to detect two or more target sequences of DNA or RNA simultaneously in a single sample is called _________ PCR.
- Conclusion
- 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: David Moffa, PhD, has over 30 years of experience in the health care industry as an executive manager, clinical laboratory director, and medical laboratory scientist. He is currently a technical consultant for Kentmere Healthcare, Wilmington, DE, and until his retirement, was the Regional Director for LabCorp, Inc. He holds a PhD in medical biochemistry from the School of Medicine, West Virginia University.
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.
