Course Title:
“DNA Barcoding and Sequencing: Species Identification and Applications”
Course Duration:
5 days (40 hours) – can be adjusted based on depth and participant background
Target Participants:
- Biology, agriculture, forestry, and environmental science students
- Laboratory technicians
- Researchers in biodiversity, forensics, and food safety
- Herbal and pharmaceutical product developers
Course Objectives:
By the end of the course, participants will be able to:
- Understand the principles of DNA barcoding and its applications.
- Extract high-quality DNA from plant, animal, or processed samples.
- Amplify barcode regions using PCR.
- Perform DNA sequencing and analyze sequence data.
- Identify species using reference databases (BOLD, GenBank).
- Apply DNA barcoding techniques in biodiversity, forensics, and herbal authentication.
Course Modules
Module 1 – Introduction to DNA Barcoding (Theory)
- Fundamentals of DNA and genes
- Concept of DNA barcoding as a “molecular ID”
- Common barcode regions: COI, rbcL, matK, ITS
- Applications in biodiversity, forensics, food safety, and research
- Overview of sequencing technologies (Sanger and NGS)
Learning Outcome: Participants can explain DNA barcoding and its role in species identification.
Module 2 – Sample Collection and DNA Extraction
- Sample types: tissue, leaves, saliva, powder
- Avoiding contamination
- DNA extraction protocols (kit-based and conventional methods)
- Assessing DNA quality (spectrophotometry, gel electrophoresis)
Hands-on Activity: Extract DNA from provided plant or animal samples.
Module 3 – PCR Amplification of Barcode Regions
- PCR principles and components
- Selection of primers for COI, rbcL, matK, ITS
- PCR setup, thermocycling, and troubleshooting
- Gel electrophoresis to verify PCR products
Hands-on Activity: Amplify barcode regions from extracted DNA.
Module 4 – DNA Sequencing
- Overview of sequencing methods (Sanger & NGS)
- Sample preparation for sequencing
- Sending PCR products for sequencing
- Understanding sequence quality and chromatograms
Hands-on Activity: Prepare PCR products for sequencing; analyze sample chromatograms.
Module 5 – Sequence Analysis and Species Identification
- Introduction to bioinformatics tools
- Aligning sequences and checking quality
- BLAST analysis in GenBank
- Barcode of Life Database (BOLD) usage
- Interpreting results for species identification
Hands-on Activity: Identify species from participant-generated sequences.
Module 6 – Applications and Case Studies
- Biodiversity monitoring and ecological research
- Herbal product authentication and quality control
- Forensic species identification
- Food fraud detection (e.g., mislabeled seafood, herbal powders)
Group Exercise: Apply DNA barcoding to real-life samples or simulated cases.
Assessment
- Practical exam: DNA extraction, PCR, gel analysis
- Report: Sequencing and species identification
- Quiz: Theory and applications of DNA barcoding
Optional Advanced Module
- Multiplex PCR and simultaneous barcoding of multiple species
- Quantitative PCR (qPCR) applications
- Metabarcoding for environmental DNA (eDNA) studies
Course Materials
- Laboratory manual (step-by-step PCR, extraction, sequencing protocols)
- Sample datasets for practice
- Reference sequences from BOLD/GenBank
- Safety and contamination prevention guide
Outcome / Certification
Participants will be certified competent in:
- Performing DNA extraction, PCR, and sequencing
- Conducting species identification using DNA barcodes
- Applying molecular techniques in research, conservation, and product verification