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Published on: August 8, 2025
Enhanced extraction efficiency of miRNA from cells by addition of Triton X-100
Eunmi Ban1, Dong-Kyu Chae, Eun Joo Song
1Molecular Recognition Research Center, Korea Institute of Science and Technology, Hwarangno 14-gil 5, Seongbuk-gu, Seoul, 136-791, South Korea.
Abstract:
The determination of microRNA (miRNA) levels in biomaterials has become important for understanding their biological functions and for the diagnosis of various diseases. An effective extraction method is needed for maximizing the recovery of miRNAs from cells, while minimizing RNA degradation during the extraction because miRNAs present only approximately 0.01 % of total RNA. In this study, we used Triton X-100 (TX-100) to improve the extraction efficiency of miRNAs with TRIzol® reagent, which is a commonly used commercial microRNA isolation kit. The concentration of TX-100 and the incubation time after the addition of TX-100 were optimized to maximize the extraction efficiency. The extraction recovery by a combination of TX-100 and TRIzol® reagent was approximately 1.9-fold greater than that by the TRIzol® reagent alone. We have established a very effective extraction method for the extraction of low-abundance miRNAs in biological samples for the determination of miRNA levels in biomaterials.
Insights
This study optimized microRNA (miRNA) extraction using Triton X-100 with TRIzol reagent. The improved method significantly enhances recovery of low-abundance miRNAs from biological samples.
Area of Science:
- Molecular Biology
- Biochemistry
Background:
- Accurate microRNA (miRNA) quantification is crucial for understanding biological roles and disease diagnostics.
- Efficient extraction is vital due to low miRNA abundance (approx. 0.01% of total RNA) and susceptibility to degradation.
Purpose of the Study:
- To develop an optimized method for enhanced microRNA extraction efficiency.
- To improve the recovery of low-abundance miRNAs from biological samples.
Main Methods:
- Utilized Triton X-100 (TX-100) in conjunction with TRIzol® reagent for microRNA isolation.
- Optimized TX-100 concentration and incubation time to maximize extraction yield.
- Compared extraction efficiency against TRIzol® reagent alone.
Main Results:
- The combination of TX-100 and TRIzol® reagent demonstrated a 1.9-fold increase in extraction recovery compared to TRIzol® alone.
- Optimized TX-100 concentration and incubation time led to maximized miRNA extraction efficiency.
Conclusions:
- A highly effective method for extracting low-abundance miRNAs has been established.
- This optimized method facilitates accurate determination of miRNA levels in biomaterials for research and diagnostics.
