Related Experiment Video
Updated: Jun 11, 2025
10:58
Profiling of Pre-micro RNAs and microRNAs using Quantitative Real-time PCR qPCR Arrays
Published on: December 3, 2010
17.3K
Evaluation of Gene Expression Profiling by QuantiGene™ 2.0 RNA Assay
Letizia Lombardelli1, Federica Logiodice2, Marie-Pierre Piccinni2
1Department of Experimental and Clinical Medicine, University of Florence, Florence, Italy. letizia.lombardelli@unifi.it.
Methods in Molecular Biology (Clifton, N.J.)
|September 30, 2024
Summary
The QuantiGene™ 2.0 technique enables multiplexed gene expression analysis to investigate immune system senescence and aging-related immune defects. This method uses branched-DNA technology for sensitive RNA detection, aiding molecular mechanism understanding.
Area of Science:
- Immunology
- Molecular Biology
- Genetics
Background:
- Aging leads to immune system senescence and defects.
- Understanding the molecular mechanisms of immune aging is crucial.
Purpose of the Study:
- To investigate the gene expression signature of immune system senescence.
- To understand the molecular mechanisms underlying immune response defects during aging.
Main Methods:
- Utilized the QuantiGene™ 2.0 technique, a multiplex platform for simultaneous RNA analysis (up to 80 targets).
- Employed branched-DNA (bDNA) technology for direct RNA transcript measurement via signal amplification.
- Assay involves cell lysis, probe hybridization to magnetic beads, signal amplification with bDNA, and fluorescent detection using R-phycoerythrin.
Main Results:
- The QuantiGene™ 2.0 technique provides specificity and multiplexing capabilities.
- Branched-DNA technology ensures high sensitivity, enabling detection of low mRNA levels.
- Fluorescence intensity, measured by Luminex, is directly proportional to target RNA levels.
Conclusions:
- QuantiGene™ 2.0 is suitable for studying gene expression in immune senescence.
- The technique facilitates understanding molecular mechanisms of immune defects in aging.
- Direct RNA measurement via signal amplification offers a sensitive approach to gene expression profiling.
Related Concept Videos
02:57Real Time RT-PCR
56.9K
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...
56.9K
03:21RNA-seq
9.8K
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...
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while...
9.8K
02:34DNA Microarrays
17.2K
Microarrays are high-throughput and relatively inexpensive assays that can be automated to analyze large quantities of data at a time. They are used in genome-wide studies to compare gene or protein expression under two varied conditions, such as healthy and diseased states. Microarrays consist of glass or silica slides on which probe molecules are covalently attached through surface functionalization. Most commonly, the slides are prepared through the chemisorption of silanes to silica...
17.2K
02:24Ribosome Profiling
3.5K
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...
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...
3.5K