Scribd
Upload
45 views12 pages

BIO 2064 Introduction and Mol. Diagnostic Applications

This document outlines the topics to be covered in a course on molecular diagnostics, including principles of PCR and RT-PCR, DNA sequencing methods, gene mutations, DNA polymorphisms, molecular detection of inherited diseases, microarrays, flow cytometry, quality assurance, and 5 laboratory practical sessions involving techniques like PCR, real-time PCR, antibody detection, methylation detection, and detecting drug resistance.

Uploaded by

gothai sivapragasam
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PPTX, PDF, TXT or read online on Scribd
45 views12 pages

BIO 2064 Introduction and Mol. Diagnostic Applications

This document outlines the topics to be covered in a course on molecular diagnostics, including principles of PCR and RT-PCR, DNA sequencing methods, gene mutations, DNA polymorphisms, molecular detection of inherited diseases, microarrays, flow cytometry, quality assurance, and 5 laboratory practical sessions involving techniques like PCR, real-time PCR, antibody detection, methylation detection, and detecting drug resistance.

Uploaded by

gothai sivapragasam
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PPTX, PDF, TXT or read online on Scribd
You are on page 1/ 12

BIO 2064: Molecular Diagnostics

• Course Outline
• Lab Practical (To Be Determined)
• Review of Molecular Biology Concepts
• Specific subject areas to be covered in molecular diagnostics
BIO 2064:Molecular Diagnostics
Course Outline
Topic

1. Principles and Application of PCR and RT-PCR

 The evolution of PCR to RT-PCR


 The application of PCR and RT-PCR
2. DNA Sequencing

 Methods and Applications in Molecular Diagnostics, Next Generation Sequencing methods


3. Gene Mutations and Mutation Detection

 Types of gene mutation and type of change within the coding sequence
 Possible consequences of gene mutation outside of the coding sequence
 Mutation rates and frequencies
 Consequences of gene mutation on protein’s structure and function
4. DNA Polymorphisms and Human Identification

 Types of molecular markers: RFLP, AFLP, VNTR, STR, SNP, STS


5. Molecular Detection of Inherited Diseases

 Nucleic acid hybridization assays, enzymatic assays, functional assays


6. Application of DNA Microarrays and Proteomics to Disease Diagnosis

 Array hybridization and gene chips


 Application of DNA and protein microarrays
 Advancement of superarray, gene expression profiling, identification of bio -markers
7. Flow Cytometry

 The principles and instrument overview


 Particle sizes comparison and the histograms produced
 Fluorescence system, cell sorting and detection
 Analysis of the plot and diagram
 Threshold, cell isolation for down-stream proceessing
8. Development of Molecular Diagnostics

 Diagnostic systems, sensitivity and limitations


 Automation and alternations for low cost solutions
 Rapid screening and instrumental analysis
9. Quality Assurance and Quality Control in the Molecular Diagnostics Laboratory
Restricted/Do Not Distribute/JT

Introduction: Biological Samples for Molecular Diagnostics

• Organ To Tissues: Methods for molecular diagnostics? (Limited)

• Tissues To Cells: Methods for molecular diagnostics?


 Flow Cytometry Method for cell analysis

• Cells To Bio-Molecules: Methods for molecular diagnostics?


 Multiple Diagnostic Tools Available, including the following molecules:

• DNA (Many Diagnostic Tools)


• RNA (Unstable, Conversion To DNA preferred)
• Proteins (Unstable, time consuming to analyze, protein tools available)
• Carbohydrates (Diagnostic tools available, not commonly used)
• Lipids (Diagnostic tools available, not commonly used)
Molecular Biology: Review
• cDNA
• Primer
• Linker
• Adapter
• cDNA library
• Types of Library
• Gene Expression Analysis
• Protein Expression Analysis
• DNA labeling
• Protein Labeling
Restricted/Do Not Distribute/JT

Molecular Diagnostics: Relevant Tools/Technologies

• DNA Methods (For Detection of DNA Polymorphism)


• DNA Methods (For Detection of DNA Mutations)
• DNA Sequencing: At Least 3 methods to sequence genes or human genome
• RNA Methods (Mainly used for gene expression analysis)
• Protein Methods (Mainly used for gene expression analysis and mutations
that change the protein function
• Cell Methods (To analyze normal and abnormal cells based on
physical properties of human cells and protein markers
Restricted/Do Not Distribute/JT

Introduction To Flow Cytometry (For Cell Analysis)


• What is Flow Cytometry?
• Components of Flow Cytometry
• How the components work?
• Collection of data through Flow Cytometry
• Applications of Flow Cytometry
Restricted/Do Not Distribute/JT

Introduction To DNA sequencing Methods

• Chemical Sequencing
• Dideoxy Sequencing
• Next Generation Sequencing (NGS)
Application of NGS (Examples)
• Whole Genome Sequencing
• Precision Medicine
• GWAS
Restricted/Do Not Distribute/JT

Introduction To Microarrays
• Types of Microarray
• How to prepare samples for microarray analysis
• Limitations/Disadvantages of microarray analysis
Quality Control and Assurance

• Sample collection and identification


• Sample quality and storage conditions
• Processing of clinical samples
• Validation of clinical tests (molecular diagnostics)
BIO 2064 Lab Practical : Molecular Diagnostics
Week Lab Practical Topics

1 Lab 1 Introduction To Molecular Diagnostics: Primer designs and PCR


amplification of EV71 DNA

2 Lab 2 Real Time PCR diagnostics of EV71

3 Lab 3 Antibody detection of EV 71 capsid protein (VP1)

4 Lab 4 Molecular detection of hyper-methylation status of DNA

5 Lab 5 Detection of Drug Resistant Cancer Cells

You might also like