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AFLP protocol comparison for microbial diversity fingerprinting
Gaia Bertani1, Maria Luisa Savo Sardaro2,3, Erasmo Neviani1
1Food and Drug Department, University of Parma, Parco Area delle Scienze 49/A, Parma, Italy.
Journal of Applied Genetics
|April 17, 2019
Summary
An improved amplified fragment length polymorphism (AFLP) protocol offers a faster, cost-effective alternative for bacterial genetic fingerprinting. This method ensures high reproducibility and resolution, aiding biodiversity and taxonomy studies.
Area of Science:
- Molecular Biology
- Microbiology
- Genetics
Background:
- Genomic DNA methods are crucial for organism identification and genotyping.
- Amplified Fragment Length Polymorphism (AFLP) is a powerful DNA fingerprinting technique with high discriminatory power and reproducibility.
- The widely used Applied Biosystems AFLP Microbial Fingerprinting Kit is no longer in production, necessitating a replacement for continuity in bacterial genetic analysis.
Purpose of the Study:
- To develop and validate an improved AFLP protocol that can serve as a reliable replacement for the discontinued commercial kit.
- To compare the performance of the improved AFLP protocol with the original commercial kit using various bacterial species.
- To ensure the continued use of previous AFLP data as a reference for future bacterial genetic fingerprinting and biodiversity studies.
Main Methods:
- An improved Amplified Fragment Length Polymorphism (AFLP) protocol was developed.
- The improved protocol was applied to bacterial strains including Listeria monocytogenes, Lactobacillus plantarum, and Streptococcus thermophilus.
- Results from the improved protocol were compared against those obtained using the previous commercial AFLP Microbial Fingerprinting Kit.
Main Results:
- The improved AFLP protocol demonstrated high reproducibility and resolution, comparable to the commercial kit.
- The new protocol was found to be faster and more cost-effective than the previous method.
- Validation confirmed the protocol's suitability for bacterial genetic fingerprinting and biodiversity studies.
Conclusions:
- The developed AFLP protocol is a viable and improved alternative to the discontinued commercial kit.
- This method supports ongoing bacterial genetic fingerprinting and biodiversity research by providing a consistent reference.
- The protocol offers enhanced efficiency and cost savings for laboratories utilizing AFLP analysis.
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