A robust lysis-based protocol for DNA extraction and long-term preservation of in situ filtered oceanic environmental samples
Abstract
The marine realm hosts very rich and diverse microbial ecosystems which remain poorly characterized due to the harsh and extreme environmental conditions for collecting them and the lack of appropriate protocol for in situ preservation of genetic material over a long time period. To address this, we developed a protocol for microbial DNA extraction and long-term preservation, with the following key features: • Lysis: The protocol uses a high-temperature lysis in a specialized lysis-preservation buffer (LP buffer), optimized to efficiently lyse microbial cells. • Preservation: It preserves DNA at 13°C for extended periods in LP buffer, ensuring stability under marine conditions. The protocol was evaluated over a 12 months period on surface Mediterranean seawater samples that were filtered on 0.22 µm PES membranes, lysed in LP buffer, and stored in it under conditions representative of sea observatories. DNA recovery, assessed by quantitative PCR targeting the bacterial 16S rRNA gene, remained constant over time, confirming the protocol efficiency for DNA preservation and its suitability for prolonged in situ preservation of microbial genetic signatures. This method provides a practical alternative to mechanical lysis and cold-chain preservation, useful for long-term monitoring of marine microbial communities through DNA metabarcoding and metagenomic analyses.
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BibTeX
@misc{roland2026robust,
title = {{A robust lysis-based protocol for DNA extraction and long-term preservation of in situ filtered oceanic environmental samples}},
author = {Agathe Roland and Alain Dolla and Valérie Chavagnac and Céline Rommevaux},
howpublished = {HAL},
year = {2026},
month = may,
url = {https://hal.science/hal-05734231v2},
note = {CC BY 4.0},
}
RIS
TY - UNPB
TI - A robust lysis-based protocol for DNA extraction and long-term preservation of in situ filtered oceanic environmental samples
AU - Agathe Roland
AU - Alain Dolla
AU - Valérie Chavagnac
AU - Céline Rommevaux
PY - 2026
DA - 2026/05/29
PB - HAL
UR - https://hal.science/hal-05734231v2
AB - The marine realm hosts very rich and diverse microbial ecosystems which remain poorly characterized due to the harsh and extreme environmental conditions for collecting them and the lack of appropriate protocol for in situ preservation of genetic material over a long time period. To address this, we developed a protocol for microbial DNA extraction and long-term preservation, with the following key features: • Lysis: The protocol uses a high-temperature lysis in a specialized lysis-preservation buffer (LP buffer), optimized to efficiently lyse microbial cells. • Preservation: It preserves DNA at 13°C for extended periods in LP buffer, ensuring stability under marine conditions. The protocol was evaluated over a 12 months period on surface Mediterranean seawater samples that were filtered on 0.22 µm PES membranes, lysed in LP buffer, and stored in it under conditions representative of sea observatories. DNA recovery, assessed by quantitative PCR targeting the bacterial 16S rRNA gene, remained constant over time, confirming the protocol efficiency for DNA preservation and its suitability for prolonged in situ preservation of microbial genetic signatures. This method provides a practical alternative to mechanical lysis and cold-chain preservation, useful for long-term monitoring of marine microbial communities through DNA metabarcoding and metagenomic analyses.
ER -