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Updated: Jul 1, 2026
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Isolation of Small Noncoding RNAs from Human Serum
Published on: June 19, 2014
Optimization of miRNA Serum Extraction: Processes and Lessons Learned in Preparation for Archival Serum Extractions
Andrew J Hoisington1,2,3, Christopher E Stamper1, Anna C Rivera4,5
1Veterans Health Administration, Rocky Mountain Mental Illness Research Education and Clinical Center (MIRECC) for Veteran Suicide Prevention, Rocky Mountain Regional Veterans Affairs Medical Center, Aurora, CO 80045, United States.
Military Medicine
|June 29, 2026
Summary
This study optimized microRNA (miRNA) extraction from aged military serum samples, enabling successful RNA sequencing for over 86% of samples. The new protocols aid in identifying early disease biomarkers and understanding toxic exposures.
Area of Science:
- Biomarker Discovery
- Epigenetics
- Occupational Health
Background:
- Military personnel face airborne hazards, potentially causing adverse health outcomes via biological mechanisms.
- Alterations in microRNA (miRNA) expression may indicate epigenetic effects of environmental or occupational exposures.
- The Department of Defense Serum Repository (DODSR) stores decades-old serum samples not ideal for RNA preservation.
Purpose of the Study:
- To develop optimized miRNA extraction protocols for RNA sequencing of aged DODSR samples.
- To establish a proof of concept for processing and sequencing over 1,000 DODSR samples.
- To provide protocols applicable to other biorepositories for analyzing stored samples.
Main Methods:
- Seven optimization experiments were performed on serum samples stored for up to 2 years.
- A total of 1,016 DODSR samples were processed using the optimized method.
- miRNA extraction and quality control were assessed via TapeStation or paired-end sequencing.
Main Results:
- Successful miRNA sequencing was achieved for over 86% of samples, with an average collection age of over 16 years.
- A significant correlation was found between sample age and successful sequencing.
- Essential optimization factors included spike RNA incorporation and controlled sample thawing using an aluminum block.
Conclusions:
- Protocols were established for measuring miRNA in DODSR samples.
- These protocols support research into early detection biomarkers.
- The findings facilitate studies on mechanistic links between occupational exposures and adverse health outcomes.
