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MicroRNA (miRNA) are short, regulatory RNA transcribed from introns—non-coding regions of a gene—or intergenic regions—stretches of DNA present between genes. Several processing steps are required to form biologically active, mature miRNA. The initial transcript, called primary miRNA (pri-mRNA), base-pairs with itself forming a stem-loop structure. Within the nucleus, an endonuclease enzyme, called Drosha, shortens the stem-loop structure into hairpin-shaped pre-miRNA. After...
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MicroRNA (miRNA) are short, regulatory RNA transcribed from introns (non-coding regions of a gene) or intergenic regions (stretches of DNA present between genes). Several processing steps are required to form biologically active, mature miRNA. The initial transcript, called primary miRNA (pri-mRNA), base-pairs with itself, forming a stem-loop structure. Within the nucleus, an endonuclease enzyme, called Drosha, shortens the stem-loop structure into hairpin-shaped pre-miRNA. After the pre-miRNA...
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Updated: Feb 5, 2026

Profiling of Pre-micro RNAs and microRNAs using Quantitative Real-time PCR qPCR Arrays
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MicroRNA Isolation from Plasma for Real-Time qPCR Array.

Isabel Witvrouwen1,2, Andreas B Gevaert1,2, Emeline M Van Craenenbroeck1,2

  • 1Laboratory of Cellular and Molecular Cardiology, Research Group Cardiovascular Diseases, Department of Translational Pathophysiological Research, University of Antwerp, Antwerp, Belgium.

Current Protocols in Human Genetics
|September 15, 2018
PubMed
Summary

Optimized plasma microRNA isolation using the mirVana PARIS kit for accurate quantification. Key recommendations include using Arabidopsis thaliana miR-159a as a spike-in control and a 100-µl elution volume.

Keywords:
arrayisolationmicroRNAplasma

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Area of Science:

  • Molecular Biology
  • Biochemistry
  • Genetics

Background:

  • MicroRNAs (miRNAs) are short non-coding RNAs regulating gene expression post-transcriptionally.
  • Plasma miRNAs are stable and considered promising biomarkers.
  • Existing isolation procedures yield variable results, necessitating optimization.

Purpose of the Study:

  • To optimize microRNA (miRNA) isolation and preamplification for plasma samples.
  • To establish a reliable method for miRNA quantification using TaqMan Low Density Arrays.
  • To provide a step-by-step protocol for researchers.

Main Methods:

  • Utilized the mirVana PARIS kit for RNA isolation from human plasma.
  • Modified the standard Thermo Fisher procedure for enhanced miRNA recovery.
  • Incorporated a preamplification step without dilution of the product.

Main Results:

  • Arabidopsis thaliana (Ath) miR-159a was identified as an effective spike-in control.
  • A 100-µl elution volume during RNA isolation maximized miRNA yield.
  • The optimized protocol ensures consistent and accurate miRNA quantification.

Conclusions:

  • The optimized protocol enhances the reliability of plasma miRNA analysis.
  • This method facilitates accurate miRNA biomarker discovery in clinical research.
  • The detailed procedure supports standardization in plasma miRNA studies.