DESS deconstructed: Is EDTA solely responsible for protection of high molecular weight DNA in this common tissue preservative?

Autor: Sarah Gayer, David Stein, Amy Sharpe, Hannah J. Appiah-Madson, Elisha Allan-Perkins, Rosalia Falco, Sonia Barrios, Daniel L. Distel
Jazyk: angličtina
Rok vydání: 2020
Předmět:
0106 biological sciences
Preservative
Gel electrophoresis of nucleic acids
Physiology
Electrophoretic techniques
DNA electrophoresis
Artificial Gene Amplification and Extension
01 natural sciences
Polymerase Chain Reaction
Biochemistry
law.invention
chemistry.chemical_compound
law
Specimen Storage
DNA extraction
Polymerase chain reaction
0303 health sciences
Multidisciplinary
Organic Compounds
Nucleic acids
Chemistry
Physiological Parameters
Physical Sciences
Medicine
Tissue Preservation
Research Article
Science
Sodium
Drug Compounding
chemistry.chemical_element
Ethylenediaminetetraacetic acid
010603 evolutionary biology
03 medical and health sciences
Extraction techniques
Genetics
Animals
Molecular Biology Techniques
Molecular Biology
Edetic Acid
030304 developmental biology
Chromatography
Ethanol
Dimethyl sulfoxide
Body Weight
Organic Chemistry
Chemical Compounds
Biology and Life Sciences
DNA
Research and analysis methods
Molecular Weight
chemistry
Specimen Preparation and Treatment
Alcohols
Storage and Handling
Preservatives
Zdroj: PLoS ONE
PLoS ONE, Vol 15, Iss 8, p e0237356 (2020)
ISSN: 1932-6203
Popis: DESS is a formulation widely used to preserve DNA in biological tissue samples. Although it contains three ingredients, dimethyl sulfoxide (DMSO), ethylenediaminetetraacetic acid (EDTA) and sodium chloride (NaCl), it is frequently referred to as a DMSO-based preservative. The effectiveness of DESS has been confirmed for a variety of taxa and tissues, however, to our knowledge, the contributions of each component of DESS to DNA preservation have not been evaluated. To address this question, we stored tissues of three aquatic taxa, Mytilus edulis (blue mussel), Faxonius virilis (virile crayfish) and Alitta virens (clam worm) in DESS, each component of DESS individually and solutions containing all combinations of two components of DESS. After storage at room temperature for intervals ranging from one day to six months, we extracted DNA from each tissue and measured the percentage of high molecular weight (HMW) DNA recovered (%R) and normalized HMW DNA yield (nY). Here, HMW DNA is defined as fragments >10 kb. For comparison, we also measured the %R and nY of HMW DNA from extracts of fresh tissues and those stored in 95% EtOH over the same time intervals. We found that in cases where DESS performed most effectively (yielding ≥ 20%R of HMW DNA), all solutions containing EDTA were as or more effective than DESS. Conversely, in cases where DESS performed more poorly, none of the six DESS-variant storage solutions provided better protection of HMW DNA than DESS. Moreover, for all taxa and storage intervals longer than one day, tissues stored in solutions containing DMSO alone, NaCl alone or DMSO and NaCl in combination resulted in %R and nY of HMW DNA significantly lower than those of fresh tissues. These results indicate that for the taxa, solutions and time intervals examined, only EDTA contributed directly to preservation of high molecular weight DNA.
Databáze: OpenAIRE