04Mini Project

DNA Barcoding & Phylogenetics

COI gene analysis of prawn & shrimp species

From the wet lab to the tree — COI barcoding and phylogenetic analysis of prawns and shrimp.

Type
Mini Project
Scope
Wet lab → Analysis
Marker
COI gene

What this project set out to solve

DNA barcoding and phylogenetic analysis of prawn and shrimp species using the COI gene span the whole molecular pipeline — from wet-lab extraction through sequencing to the trees that reveal species relationships.

A reproducible barcode pipeline needs each link in the chain: extraction, amplification, sequencing, identification, alignment and phylogenetics.

How it was approached

Performed DNA extraction, PCR amplification of the mitochondrial COI gene, agarose gel electrophoresis and Sanger sequencing for species identification.

Conducted NCBI BLAST-based species identification and Multiple Sequence Alignment (MSA) using Clustal Omega.

Generated DNA barcodes and constructed phylogenetic trees to evaluate genetic relationships among the identified species.

Workflow & architecture

Barcoding pipeline

  1. DNA extractionWet lab

    DNA extraction was performed for prawn and shrimp species identification.

  2. COI PCRMitochondrial COI

    PCR amplification of the mitochondrial COI gene was carried out.

  3. Gel & SangerSequencing

    Agarose gel electrophoresis and Sanger sequencing were performed.

  4. Species IDNCBI BLAST

    NCBI BLAST-based species identification was conducted.

  5. AlignmentClustal Omega

    Multiple Sequence Alignment (MSA) was performed using Clustal Omega.

  6. PhylogeneticsMEGA

    Phylogenetic analysis was carried out using MEGA.

  7. Barcodes & treeBOLD Systems

    DNA barcodes were generated and phylogenetic trees constructed to evaluate genetic relationships among identified prawn and shrimp species.

Stage 01 / 07

DNA extraction

Wet lab

DNA extraction was performed for prawn and shrimp species identification.

  • Molecular lab — DNA extraction, COI PCR, agarose gel electrophoresis, Sanger sequencing.
  • Identification — NCBI BLAST-based species identification, with BOLD Systems as reference.
  • Alignment — Multiple Sequence Alignment (MSA) with Clustal Omega.
  • Phylogenetics — tree construction and analysis with MEGA.

What it was built with

Wet lab

DNA extractionCOI PCRAgarose gel electrophoresisSanger sequencing

Identification

NCBI BLASTBOLD Systems

Alignment

Clustal OmegaMultiple Sequence Alignment (MSA)

Phylogenetics

MEGA

What was built

  • DNA extraction followed by PCR amplification of the mitochondrial COI gene, agarose gel electrophoresis and Sanger sequencing.
  • NCBI BLAST-based species identification and Multiple Sequence Alignment (MSA) using Clustal Omega.
  • Phylogenetic analysis using MEGA, with DNA barcodes generated and trees constructed for the identified species.

Results & output

Results and detailed analysis available in the project repository.

What it taught me

  • A species barcode project is a complete pipeline — wet-lab work, sequencing and analysis each matter for a trustworthy identification.
  • Standard, reproducible links — BLAST, Clustal Omega, MEGA — connect molecular data to phylogenetic inference.
  • The COI marker ties molecular taxonomy to evolutionary relationships in a single workflow.
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