Enhanced extraction efficiency of miRNA from cells by addition of Triton X-100

Eunmi Ban; Dong Kyu Chae; Eun Joo Song

doi:10.1007/s00216-013-7170-0

Enhanced extraction efficiency of miRNA from cells by addition of Triton X-100

Eunmi Ban, Dong Kyu Chae, Eun Joo Song

Department of Pharmaceutical Sciences

Research output: Contribution to journal › Article › peer-review

7 Scopus citations

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.

Original language	English
Pages (from-to)	7535-7539
Number of pages	5
Journal	Analytical and Bioanalytical Chemistry
Volume	405
Issue number	23
DOIs	https://doi.org/10.1007/s00216-013-7170-0
State	Published - Sep 2013

Bibliographical note

Funding Information:
Acknowledgments We gratefully acknowledge the support of GRL Program (20110021713) from the National Research Foundation of Korea and an institutional grant from the Korea Institute of Science and Technology (2E23810 and 2E24080).

Keywords

Extraction efficiency
miRNA
Triton X-100

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10.1007/s00216-013-7170-0

Cite this

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title = "Enhanced extraction efficiency of miRNA from cells by addition of Triton X-100",

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{\textregistered} 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{\textregistered} reagent was approximately 1.9-fold greater than that by the TRIzol{\textregistered} 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.",

keywords = "Extraction efficiency, miRNA, Triton X-100",

author = "Eunmi Ban and Chae, {Dong Kyu} and Song, {Eun Joo}",

note = "Funding Information: Acknowledgments We gratefully acknowledge the support of GRL Program (20110021713) from the National Research Foundation of Korea and an institutional grant from the Korea Institute of Science and Technology (2E23810 and 2E24080).",

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AU - Ban, Eunmi

AU - Chae, Dong Kyu

AU - Song, Eun Joo

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N2 - 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.

AB - 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.

KW - Extraction efficiency

KW - miRNA

KW - Triton X-100

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JO - Analytical and Bioanalytical Chemistry

JF - Analytical and Bioanalytical Chemistry

IS - 23

ER -

Enhanced extraction efficiency of miRNA from cells by addition of Triton X-100

Abstract

Bibliographical note

Keywords

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