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Laboratory Protocol for Genetic Gut Content Analyses of Aquatic Macroinvertebrates Using Group-specific rDNA Primers

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Using microwaves to prepare gastropods for DNA barcoding.

L A Galindo¹, N Puillandre, E E Strong

¹Muséum National d'Histoire Naturelle, Département Systématique et Evolution, ISyEB Institut (UMR 7205 CNRS/UPMC/MNHN/EPHE), 43, Rue Cuvier, 75231, Paris, France.

Molecular Ecology Resources

|January 24, 2014

Summary

This summary is machine-generated.

Microwaving gastropods quickly separates animals from shells, preserving DNA and shells. This method significantly increases DNA extraction success and productivity for molluscan research.

Keywords:

DNA damage PCR optimization shelled molluscs tissue preparation voucher

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Area of Science:

Marine Biology
Genetics
Molecular Biology

Background:

DNA extraction from gastropods is challenging due to shell retraction, hindering ethanol penetration and tissue fixation.
Existing methods like chemical relaxation, drilling, or boiling are time-consuming, pose safety risks, and limit large-scale fieldwork.
Preserving shell vouchers is crucial for linking genetic data with taxonomic identity.

Purpose of the Study:

To evaluate a novel microwave (MW) method for rapidly separating gastropods from their shells for DNA extraction.
To compare the efficacy of the MW method against traditional anesthetizing/drilling techniques.
To assess the impact of the MW method on DNA quality, quantity, and subsequent sequencing.

Main Methods:

Live gastropods were placed in a microwave (MW) oven to rapidly heat internal water, causing muscle separation from the shell.
The intact animal body was removed, and the shell voucher was preserved undamaged.
DNA was extracted and amplified using PCR from MW-treated specimens and compared to those treated with anesthetizing/drilling.

Main Results:

Microwaving effectively separated gastropod bodies from shells without damaging the shell voucher.
DNA quantity and quality were comparable between MW-treated and traditionally treated specimens.
Sequencing results were identical within species, confirming the method's reliability for genetic analysis.
Large-scale field implementation showed a higher percentage of DNA extraction success.

Conclusions:

The microwave method offers a rapid, safe, and effective alternative for preparing gastropods and other molluscs for DNA extraction.
This technique significantly increases productivity (three- to fivefold) in fieldwork compared to conventional methods.
The MW method preserves both genetic integrity and valuable shell vouchers, streamlining molecular and taxonomic studies.