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
Abstract
Sample preparation optimization by sodium acetate precipitation for DNA extraction from Dry-Sponge samples.
Sample Preparation
Sample Preparation
15h 55m
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.