Accelerated and efficient method for isolating microRNA from human blood plasma
https://doi.org/.org/10.21285/2227-2925-2019-9-1-53-59
Abstract
He study is dedicated to solving the problem of microRNA extraction from human blood for further use of the microRNA profile in the diagnosis of various diseases. The aim of the work is to choose an effective, accelerated method for the isolation of microRNA from human blood plasma, sufficient in quantity for diagnosing in clinical practice. Comparison of three methods for obtaining microRNA was carried out: 1) using a silicon oxide-based QIAamp RNA Blood Mini Kit column, 2) phenol-chloroform extraction using TRIzol LS Reagent and 3) phenol-chloroform extraction using TRIzol Reagent. It was established that for the most complete isolation of total human RNA from human blood plasma in order to obtain microRNA, TRIzol LS Reagent can be efficiently used. Because its use allows obtaining microRNA in less time - 30-40 min, and in an amount sufficient for clinical analysis (total RNA - up to 0,116 ng/μl, and microRNA - 350,3 picograms/μl), and also with the least amount of concomitant messenger RNA and long non-coding RNA. Using TRIzol LS Reagent allows you to increase the output of miRNA by 2,7 times compared with the use of TRIzol Reagent.
About the Authors
I. A. Letova
Kazan National Research Technological University
Russian Federation
Russian Federation
S. A. Madumarov
Kazan Federal University
Russian Federation
Russian Federation
M. A. Sysoyeva
Kazan National Research Technological University
Russian Federation
Russian Federation
R. Z. Shah Mahmud
Kazan Federal University; Interdisciplinary Center for Proteomic Research Institute of Fundamental Medicine and Biology Kazan Federal University e-mail: raihan.shah@kpfu.ru
Russian Federation
Russian Federation
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Review
For citations:
Letova I.A., Madumarov S.A., Sysoyeva M.A., Shah Mahmud R.Z. Accelerated and efficient method for isolating microRNA from human blood plasma. Proceedings of Universities. Applied Chemistry and Biotechnology. 2019;9(1):53-59. (In Russ.) https://doi.org/.org/10.21285/2227-2925-2019-9-1-53-59
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