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Updated: Oct 31, 2025
Isolation of Small Noncoding RNAs from Human Serum
Published on: June 19, 2014
Evaluation of five column-based isolation kits and their ability to extract miRNA from human milk
Emelie Ahlberg1, Maria C Jenmalm1, Lina Tingö1,2
1Division of Inflammation and Infection, Department of Biomedical and Clinical Sciences, Linköping University, Linköping, Sweden.
Abstract:
MicroRNA can be found in various body fluids, including breast milk. MicroRNA may be transferred from mother to infant via breast milk and potentially regulate the development of the infant's immune system on a post-transcriptional level. This study aimed to determine the microRNA extraction efficiency of five RNA extraction kits from human skim milk samples. Their efficiency was determined by comparing microRNA concentrations, total RNA yield and purity. Furthermore, hsa-miR-148a-3p expression and the recovery of an exogenous control, cel-miR-39-3p, were quantified using qPCR. Each kit extracted different amounts of microRNA and total RNA, with one kit tending to isolate the highest amount of both RNA species. Based on these results, the extraction kit ReliaPrep™ miRNA Cell and Tissue Miniprep System from Promega was found to be the most appropriate kit for microRNA extraction from human skim milk. Moreover, further research is needed to establish a standardized protocol for microRNA extraction from breast milk.
Insights
This study evaluated RNA extraction kits for isolating microRNA from human breast milk. The Promega ReliaPrep™ system demonstrated the highest efficiency for microRNA and total RNA extraction.
Area of Science:
- Biochemistry
- Molecular Biology
- Neonatal Immunology
Background:
- MicroRNAs (miRNAs) are present in human breast milk.
- Maternal miRNAs in milk may influence infant immune system development post-transcriptionally.
- Standardized methods for miRNA extraction from breast milk are lacking.
Purpose of the Study:
- To assess the efficiency of five RNA extraction kits for isolating miRNA from human skim milk.
- To compare miRNA yield, total RNA yield, and purity across different kits.
- To identify the most suitable kit for reliable miRNA extraction from breast milk.
Main Methods:
- Human skim milk samples were used for RNA extraction.
- Five commercial RNA extraction kits were evaluated.
- Quantification of miRNA and total RNA yield, purity, and specific miRNA (hsa-miR-148a-3p) expression via qPCR.
- Recovery of exogenous control (cel-miR-39-3p) was assessed.
Main Results:
- Significant variations in miRNA and total RNA extraction yields were observed among the kits.
- One kit consistently yielded higher amounts of both RNA species.
- The Promega ReliaPrep™ miRNA Cell and Tissue Miniprep System showed superior performance.
- qPCR confirmed differential expression and recovery rates influenced by the extraction kit.
Conclusions:
- The Promega ReliaPrep™ miRNA Cell and Tissue Miniprep System is recommended for miRNA extraction from human skim milk.
- Further research is necessary to develop a standardized protocol for miRNA isolation from breast milk.
- Optimized extraction is crucial for studying the role of milk miRNAs in infant health.
