Apr 22, 2025
Dry-Sponge DNA extraction
- Amir reza Varzandi1
- 1University of Turin
Protocol Citation: Amir reza Varzandi 2025. Dry-Sponge DNA extraction. protocols.io https://dx.doi.org/10.17504/protocols.io.e6nvwew77vmk/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: In development
We are still developing and optimizing this protocol
Created: April 22, 2025
Last Modified: April 22, 2025
Protocol Integer ID: 137538
Keywords: sponge dna extraction sample preparation optimization, dna extraction from dry, dna extraction, sponge sample, extraction, sodium acetate precipitation, sample preparation
Abstract
Sample preparation optimization by sodium acetate precipitation for DNA extraction from Dry-Sponge samples.
Troubleshooting
Sample Preparation
15h 55m
Extract the sponge liquid by squizing the sponge.
5m
Move (pour) the liquid content extracted from the sponge into 50 mL tubes (expect 15 mL eg. Longmire's buffer)
5m
Add 33 mL of pure ethanol and 1.m mL of Sodium Acetate 3M to the sample tube (achieving approx. 50 mL of liquid voulme).
5m
incubate the samples overnight at -20 °C
12h
Cool down the centrifuge to 4-8 °C
15m
Centrifuge at 4000g for 80 min at 6°C.
1h 20m
Discard the supernatant and take up to 250 mg of the sediment into PowerBead tubes (Qiagen, Hilden, Germany)
5m
Follow the DNeasy PowerSoil kit instruction for the DNA extraction.
2h