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Published on: October 26, 2017
miRNA Stability in Frozen Plasma Samples
Francesca Balzano1, Marta Deiana2, Silvia Dei Giudici3
1Department of Biomedical Sciences, University of Sassari, vl. San Pietro 43b, Sassari 07100, Italy. mariafrancesca22@virgilio.it.
Abstract:
MicroRNAs (miRNAs) represent a family of small non-coding ribonucleic acids that post-transcriptionally inhibits the expression of their target messenger RNAs (mRNAs), thereby acting as general gene repressors. In this study we examined the relative quantity and stability of miRNA subjected to a long period of freezing; we compared the stability of eight miRNAs in the plasma of five human healthy controls before freezing and after six and 12 months of storage at -80 °C. In addition, we examined the plasma frozen for 14 years and the amount of miRNA still available. Using a Life Technologies protocol to amplify and quantify plasma miRNAs from EDTA (Ethylene Diamine Tetraacetic Acid)-treated blood, we analyzed the stability of eight miRNAs, (miR-125b-5p, miR-425-5p, miR-200b-5p, miR-200c-3p, miR-579-3p, miR-212-3p, miR-126-3p, and miR-21-5p). The miRNAs analyzed showed a high stability and long frozen half-life.
Insights
MicroRNAs (miRNAs) are stable when frozen for extended periods. This study found that plasma miRNAs remained quantifiable after 14 years of frozen storage, indicating their long-term stability.
Area of Science:
- Molecular Biology
- Genetics
- Biochemistry
Background:
- MicroRNAs (miRNAs) are small non-coding RNAs that regulate gene expression post-transcriptionally.
- They play crucial roles in various biological processes and diseases.
- Assessing miRNA stability in biological samples is vital for accurate research and diagnostics.
Purpose of the Study:
- To evaluate the stability and quantity of specific plasma miRNAs after prolonged frozen storage.
- To determine the long-term viability of miRNAs in samples stored for up to 14 years at -80 °C.
Main Methods:
- Eight specific miRNAs were quantified in plasma samples from healthy controls.
- Samples were analyzed before freezing and after storage periods of 6, 12 months, and 14 years at -80 °C.
- A Life Technologies protocol was used for miRNA amplification and quantification from EDTA-treated blood.
Main Results:
- The analyzed miRNAs demonstrated high stability over extended frozen storage periods.
- Quantifiable amounts of miRNAs were still present in plasma samples frozen for 14 years.
- The study indicates a long frozen half-life for the investigated plasma miRNAs.
Conclusions:
- Plasma miRNAs are highly stable under long-term frozen storage conditions (-80 °C).
- This stability supports the use of frozen plasma samples for miRNA research and biomarker discovery.
- The findings validate the reliability of miRNA analysis in archived biological specimens.
