Related Experiment Video
Updated: May 27, 2026
07:27
High Throughput MicroRNA Profiling: Optimized Multiplex qRT-PCR at Nanoliter Scale on the Fluidigm Dynamic ArrayTM IFCs
Published on: August 3, 2011
Whole-genome RT-qPCR microRNA expression profiling.
Pieter Mestdagh1, Stefaan Derveaux, Jo Vandesompele
1Center for Medical Genetics, Ghent University, De Pintelaan 185, Ghent 9000, Belgium.
Methods in Molecular Biology (Clifton, N.J.)
|December 2, 2011
Summary
Accurate quantification of microRNAs (miRNAs), small gene-regulating molecules, is challenging due to their size. This chapter details RT-qPCR technologies for whole genome miRNA expression analysis.
Area of Science:
- Molecular Biology
- Genetics
- Biochemistry
Background:
- MicroRNAs (miRNAs) are small noncoding RNA molecules regulating gene expression.
- Dysregulated miRNA expression is linked to various human diseases, including cancer.
- The small size of miRNAs (20-22 nucleotides) poses challenges for accurate quantification.
Purpose of the Study:
- To present various RT-qPCR technologies for miRNA expression quantification.
- To address the challenges in accurately measuring miRNA levels.
- To enable whole genome miRNA expression analysis.
Main Methods:
- Review of different Reverse Transcription quantitative Polymerase Chain Reaction (RT-qPCR) technologies.
- Focus on methods suitable for small RNA quantification.
- Discussion of techniques for whole genome miRNA profiling.
Main Results:
- RT-qPCR technologies offer viable solutions for miRNA quantification.
- Specific methods are highlighted for their suitability in analyzing small RNA expression.
- The presented technologies facilitate comprehensive miRNA profiling.
Conclusions:
- RT-qPCR is a powerful tool for accurate miRNA expression analysis.
- Addressing quantification challenges is crucial for understanding miRNA's role in disease.
- Whole genome miRNA expression profiling is achievable with advanced RT-qPCR techniques.
Related Concept Videos
01:22MicroRNAs
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 ends...
02:57Real Time RT-PCR
Real-time reverse transcription-polymerase chain reaction, or Real-time RT-PCR, is an analytical tool used to determine the expression level of target genes. The method involves converting mRNA to complementary DNA with the help of an enzyme known as reverse transcriptase, followed by the PCR amplification of the cDNA. These two processes can be performed simultaneously in a single tube or separately as a two-step reaction.
The real-time quantification of the number of amplified products is...
The real-time quantification of the number of amplified products is...
02:24Ribosome Profiling
Ribosome profiling or ribo-sequencing is a deep sequencing technique that produces a snapshot of active translation in a cell. It selectively sequences the mRNAs protected by ribosomes to get an insight into a cell’s translation landscape at any given point in time.
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique helps...
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique helps...