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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...
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...

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Single-cell Gene Expression Profiling Using FACS and qPCR with Internal Standards
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Published on: February 25, 2017

Methods for quantitation of gene expression.

Vigdis Nygaard1, Eivind Hovig

  • 1Department of Tumor Biology, Institute for Cancer Research, Norwegian Radium Hospital, 0310 Oslo, Norway.

Frontiers in Bioscience (Landmark Edition)
|March 11, 2009
PubMed
Summary
This summary is machine-generated.

Quantitative gene expression analysis uses various methods, each with unique sensitivities and limitations. Method selection depends on study goals, especially when measuring low mRNA levels or non-coding transcripts.

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Area of Science:

  • Molecular Biology
  • Genomics
  • Transcriptomics

Background:

  • Gene expression is quantitatively measured at the transcriptional level using diverse low- to high-throughput methods.
  • Method sensitivity varies based on technology and target mRNA quantity, leading to potential discrepancies between platforms.
  • The complex transcriptome necessitates careful method selection based on specific study objectives, considering inherent advantages and disadvantages.

Purpose of the Study:

  • To review quantitative gene expression measurement methods.
  • To highlight the impact of sample size and stochasticity on measurement accuracy.
  • To discuss the role of microarray technology in analyzing non-coding transcripts.

Main Methods:

  • Review of various gene expression measurement technologies.
  • Discussion of strategies like global mRNA amplification for small samples.
  • Analysis of microarray technology's application in transcriptomics.

Main Results:

  • Method choice is critical due to variable correlations between platforms and sensitivity limitations.
  • Stochastic events pose challenges for accurate quantification at extremely low input levels.
  • Microarray technology remains valuable for non-coding transcript analysis.

Conclusions:

  • Gene expression analysis requires careful consideration of method-specific limitations.
  • Advancements in technology may further refine transcriptomic studies.
  • Microarrays offer a versatile approach for querying non-coding RNA levels.