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Related Concept Videos

Real Time RT-PCR02:57

Real 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...
Ribosome Profiling02:24

Ribosome 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...
RNA-seq03:21

RNA-seq

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 microarray-based...
DNA Microarrays02:34

DNA Microarrays

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...
Regulation of Expression at Multiple Steps01:23

Regulation of Expression at Multiple Steps

The gene expression in cells is regulated at different stages: (i) transcription, (ii) RNA processing, (iii) RNA localization, and (iv) translation. Transcriptional regulation is mediated by regulatory proteins such as transcription factors, activators, or repressors—these control gene expression by initiating or inhibiting the transcription of genes. Once a precursor or pre-mRNA is produced, it undergoes post-transcriptional modification, including 5' capping, splicing, and the addition of a...
mRNA Stability and Gene Expression02:51

mRNA Stability and Gene Expression

The structure and stability of mRNA molecules regulates gene expression, as mRNAs are a key step in the pathway from gene to protein. In eukaryotes, the half-life of mRNA varies from a few minutes up to several days. mRNA stability is essential in growth and development. The absence of the proteins regulating its stability, such as tristetraprolin in mice, can cause systemic issues, including bone marrow overgrowth, inflammation, and autoimmunity.
Cis-acting Elements involved in mRNA stability

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Related Experiment Video

Updated: May 30, 2026

Absolute Quantification of Plasma MicroRNA Levels in Cynomolgus Monkeys, Using Quantitative Real-time Reverse Transcription PCR
10:23

Absolute Quantification of Plasma MicroRNA Levels in Cynomolgus Monkeys, Using Quantitative Real-time Reverse Transcription PCR

Published on: February 12, 2018

Absolute and relative quantification of mRNA expression (transcript analysis).

Andrew Hayes1, Bharat M Rash, Leo A H Zeef

  • 1Faculty of Life Sciences, The University of Manchester, Manchester, UK. andy.hayes@manchester.ac.uk

Methods in Molecular Biology (Clifton, N.J.)
|August 25, 2011
PubMed
Summary
This summary is machine-generated.

This protocol details yeast messenger RNA (mRNA) expression analysis using Affymetrix GeneChip® and quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR). It offers a flexible, inexpensive, and sensitive method for transcriptomics data validation.

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Profiling of Pre-micro RNAs and microRNAs using Quantitative Real-time PCR (qPCR) Arrays

Published on: December 3, 2010

Area of Science:

  • Molecular Biology
  • Genomics
  • Biotechnology

Background:

  • Transcriptomics enables large-scale analysis of gene expression.
  • Quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) is a standard method for gene expression validation.
  • Affymetrix GeneChip® technology provides a platform for high-throughput transcriptomics.

Purpose of the Study:

  • To describe a comprehensive pipeline for transcript analysis in yeast.
  • To provide a detailed protocol for validating transcriptomics data using qRT-PCR.
  • To offer a flexible, inexpensive, sensitive, and user-friendly approach.

Main Methods:

  • Utilizing Affymetrix GeneChip® for initial transcriptomics data generation.
  • Implementing quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) for data validation.
  • Outlining necessary hardware, software, and reagents for the protocol.

Main Results:

  • A validated pipeline for yeast mRNA expression analysis is presented.
  • The protocol facilitates accurate quantification of mRNA levels.
  • The described method integrates two key transcriptomics techniques.

Conclusions:

  • This protocol offers a robust method for yeast transcriptomics research.
  • The described approach is suitable for researchers seeking cost-effective and sensitive gene expression analysis.
  • The pipeline ensures reliable validation of transcriptomics data.