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Protocol for mitochondrial DNA isolation from Macrobrachium rosenbergii and Penaeus monodon using differential
Dipta Chandra Pal1, Muhammad Manjurul Karim1
1Department of Microbiology, University of Dhaka, Dhaka 1000, Bangladesh.
STAR Protocols
|June 3, 2025
Summary
This study details a kit-free protocol for extracting mitochondrial DNA from shrimp and prawns. The method uses differential centrifugation for applications in population genetics and evolutionary biology.
Area of Science:
- Molecular Biology
- Genetics
- Evolutionary Biology
Background:
- The mitochondrial genome (mitogenome) is crucial for molecular phylogenetics, evolutionary biology, and population genetics.
- Existing methods for mitogenome extraction often rely on commercial kits.
Purpose of the Study:
- To present a cost-effective, kit-free protocol for extracting mitochondrial DNA from shrimp (Macrobrachium rosenbergii) and prawn (Penaeus monodon).
- To provide detailed steps for sample preparation, mitochondrial isolation, and genome sequencing.
Main Methods:
- Differential centrifugation for crude mitochondrial extraction.
- Mitochondrial DNA isolation and purity assessment.
- Confirmation of mitogenome presence and sequencing procedures.
Main Results:
- A comprehensive protocol for isolating and sequencing the mitochondrial genome from shrimp and prawn species was successfully developed.
- The protocol avoids the need for commercial extraction kits, offering a more accessible approach.
Conclusions:
- The presented protocol offers a viable, kit-free alternative for researchers studying mitochondrial genomes in crustaceans.
- This method supports advancements in molecular phylogenetics, population genetics, and evolutionary studies.
