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Hospital virgen del rocío

A sustainable alternative for 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.

Objective

Compare the quality and functionality of plasma samples stabilised and stored at room temperature versus frozen at -80ºC, with the aim of extracting circulating DNA after a period of storage at room temperature.

Materials and Methods

Samples used for the study

Plasma samples

30 samples

Frozen

Procedure

The analytical study will be carried out on remnants of clinical or research samples from patients with primary or metastatic lung or colon cancer. These samples have already been diagnosed, and the presence of any molecular alterations (mainly mutations) can be rechecked.

The frozen plasma samples will be thawed and divided into two. One part will be refrozen at -80°C, while the other will be freeze-dried using 300K Solutions technology and stored at room temperature. After 3 months under desiccated conditions, freeze-dried samples are re-evaluated and directly compared with their corresponding frozen according to the QC established in the following section.

Quality Assurance

DNA Concentration

DNA integrity number

EGFR, RAS and B-RAF gene mutations

All samples processed and stored under the different conditions (freeze-dried or frozen) will be analyzed simultaneously to minimize inter-assay variability. The following techniques will be used for quality assessment:
  • Quantification of double-stranded DNA using the Qubit® fluorometric quantification system.
  • Assessment of DNA integrity using the Genomic DNA (gDNA) ScreenTape assay on the TapeStation system.
  • EGFR, RAS and B-RAF gene mutations by PCR (Frozen samples underwent PCR testing at diagnosis to detect mutations in genes such as EGFR, RAS and BRAF, depending on whether the cancer was in the lungs or colon. The same PCR test will be repeated after lyophilisation to check for any changes.)

Results

Coming soon… 

Conclusion

Coming soon… 

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