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Related Concept Videos

RNA-seq03:21

RNA-seq

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RNA sequencing, or RNA-Seq, is a high-throughput sequencing technology used to study the transcriptome of a cell. Transcriptomics helps to interpret the functional elements of a genome and identify the molecular constituents of an organism. Additionally, it also helps in understanding the development of an organism and the occurrence of diseases. 
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while...
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MicroRNAs01:22

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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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MicroRNAs01:22

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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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Related Experiment Video

Updated: Feb 21, 2026

A Complete Pipeline for Isolating and Sequencing MicroRNAs, and Analyzing Them Using Open Source Tools
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MicroRNA Expression Analysis Using Small RNA Sequencing Discovery and RT-qPCR-Based Validation.

Alan Van Goethem1, Pieter Mestdagh1, Tom Van Maerken1

  • 1Center for Medical Genetics, Ghent University, Ghent, Belgium.

Methods in Molecular Biology (Clifton, N.J.)
|October 8, 2017
PubMed
Summary

Accurate quantification of microRNAs (miRNAs), crucial gene expression regulators implicated in diseases like cancer, is challenging. This chapter details two key technologies for precise miRNA measurement: small RNA sequencing and RT-qPCR.

Keywords:
NormalizationRT-qPCRSmall RNA sequencingmiRNAmiRNA annotation

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • MicroRNAs (miRNAs) are small noncoding RNA molecules regulating gene expression.
  • Aberrant miRNA expression is linked to various diseases, notably cancer.
  • Accurate miRNA quantification presents technical challenges due to their small size and sequence homology.

Purpose of the Study:

  • To present and compare two distinct technologies for miRNA quantification.
  • To provide insights into the methodologies for accurate miRNA expression analysis.

Main Methods:

  • Small RNA sequencing: A high-throughput method for comprehensive miRNA profiling.
  • Reverse Transcription Quantitative Polymerase Chain Reaction (RT-qPCR): A sensitive and specific method for targeted miRNA detection.

Main Results:

  • Both small RNA sequencing and RT-qPCR offer distinct advantages for miRNA quantification.
  • The choice of technology depends on specific research needs, such as comprehensive profiling versus targeted analysis.

Conclusions:

  • Accurate miRNA quantification is essential for understanding their role in disease.
  • Small RNA sequencing and RT-qPCR are valuable tools for researchers studying miRNA expression and its implications in various biological contexts, including cancer.