Fundació per al Foment de la Investigació Sanitària i Biomèdica de la Comunitat Valenciana (FISABIO)
A sustainable alternative to deep-freezing for DNA and plasma preservation.
DNA is a naturally stable molecule that is well suited to long-term storage. However, inadequate storage makes it susceptible to degradation processes that can compromise its quality—especially when it comes to meeting the demands of new and more sensitive molecular biology techniques.
As a result, despite its supposed stability, DNA is still routinely stored in ultra-low temperature freezers, occupying between 10% and 20% of available storage capacity in biobanks and laboratories.
Plasma, on the other hand, is a complex biological fluid with a high protein content and significant clinical relevance. It is particularly sensitive to freeze-thaw cycles and storage conditions, which can affect the stability of its components and compromise downstream analyses.
Objective
To assess the technical feasibility of 300K Solutions technology for the dry stabilization DNA. The study aims to compare the quality and functionality of DNA extracted from human blood and freeze-dried using 300K Solutions’ technology with samples traditionally stored at -80°C. This validation is part of a study into blood biomarkers for the diagnosis of Alzheimer’s disease.
Materials and Methods
Samples used for the study
30 DNA samples
extracted from peripheral blood
20 DNA samples
frozen at -80 ºC for years
Procedure
Two types of samples were used for this validation: DNA extracted from fresh whole blood and DNA previously stored at -80°C, surplus from previous projects.
For new DNA samples obtained from whole blood using the Durviz kit, half of the aliquots were stored conventionally at -80ºC and half were freeze-dried using 300K technology. Before storage, all samples were assessed by quality control (nanodrop and gel electrophoresis) to ensure that they met purity and concentration standards. Freeze-dried samples were stored at room temperature and analysed at one and six months to compare integrity with frozen aliquots.
Previously stored frozen samples were thawed and then subjected to the same lyophilisation process as new samples. Due to reduced availability, the analysis focused on a single point of assessment at three months post freeze-drying. After rehydration, quality controls were performed to confirm DNA integrity.
Quality Assurance
Ratio A260/280
DNA concentration and purity
Electrophoresis
DNA Integrity
Bioanalyser
Fragment size
Genetic analysis
blood biomarkers for Alzheimer's disease.
All samples were subjected to quality control before and after lyophilization. Concentration and purity were assessed by Nanodrop, integrity by gel electrophoresis and fragment size by Bioanalyser. For new samples, these tests were performed before storage and repeated after 1 and 6 months. For previously stored samples, they were assessed after rehydration, three months after lyophilization. Genetic analyses were also included in the study of blood biomarkers for Alzheimer’s disease.
Results
Coming soon…
Conclusion
Coming soon…
