Jun 22, 2024

Public workspaceDESS (DMSO/EDTA/NACL) Protocol

DOI
https://dx.doi.org/10.17504/protocols.io.5jyl822n9l2w/v1
  • Mirayana Marcelino Barros1,
  • Tiago Pereira2,
  • Alejandro De Santiago Perez1,
  • Hunter Powell3,
  • Jenna Brown4
  • 1University of Georgia;
  • 2Department of Marine Sciences, University of Georgia;
  • 3UGA;
  • 4University of Georgia- Holly Bik
  • Bik Lab UGA
Jenna Brown
Icon indicating open access to content
QR code linking to this content
DOI: https://dx.doi.org/10.17504/protocols.io.5jyl822n9l2w/v1
Protocol Citation: Mirayana Marcelino Barros, Tiago Pereira, Alejandro De Santiago Perez, Hunter Powell, Jenna Brown 2024. DESS (DMSO/EDTA/NACL) Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.5jyl822n9l2w/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: Working
We use this protocol and it's working
Created: May 29, 2024
Last Modified: June 22, 2024
Protocol Integer ID: 100828
Keywords: protocol dess, dess, dmso, protocol, edta, nacl, solution for the preservation, sample, preservation
Funders Acknowledgements:
Holly Bik
Abstract
DESS (DMSO/EDTA/NACL) protocol instructing how to make the solution for the preservation of samples.
Make 2L stock solution of 0.5M EDTA; if stock is already made, proceed to step 4.
Combine Amount372.24 g disodium EDTA and Amount500 mL deionized water.
Tip: Disodium EDTA is labeled PINK in the Bik Lab.
Add enough 5M NaOH to the solution to bring it to a pH of Ph8.0
Tip: NaOH is labeled GREEN in the Bik Lab.
Tip: In some cases, this can be as much as 500 mL. The EDTA will begin to dissolve around a pH of Ph7.0 .
Bring final volume to Amount2 L with deionized water once all disodium EDTA has dissolved.
Prepare 2L DESS from 0.5M EDTA.
Combine Amount1 L 0.5M disodium EDTA, Amount400 mL Dimethyl Sulfoxide/DMSO, and Amount600 mL deionized water in a 4000 mL Erlenmeyer flask.
Tip: 0.5M disodium EDTA is labeled YELLOW and Dimethyl Sulfoxide/DMSO is labeled ORANGE in the Bik Lab.
Put magnetic stir bars in the solution and place flask onto a hot plate. Turn the magnetic stirring function on and leave it at room temperature.
Add enough NaCl to saturate the solution: Add NaCl in 100g increments until 300g is reached, and then proceed to adding smaller amounts until salt is no longer dissolving. Solution should saturate with the addition of ~300-400g NaCl.
Tip: NaCl is labeled RED in the Bik Lab.
Tip: This step can take hours: BE PATIENT. There may be leftover salt residue once the solution is fully saturated.