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Related Concept Videos

Real Time RT-PCR02:57

Real Time RT-PCR

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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...
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Cell Specific Gene Expression01:58

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Multicellular organisms contain a variety of structurally and functionally distinct cell types, but the DNA in all the cells originated from the same parent cells. The differences in the cells can be attributed to the differential gene expression. Liver cells, whose functions include detoxification of blood, production of bile to metabolize fats, and synthesis of proteins essential for metabolism, must express a specific set of genes to perform their functions. Gene expression also varies with...
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Cell Specific Gene Expression01:58

Cell Specific Gene Expression

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What is Gene Expression?01:42

What is Gene Expression?

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Overview
Gene expression is the process in which DNA directs the synthesis of functional products, that is, proteins. Cells can regulate gene expression at various stages. It allows organisms to generate different cell types and enables cells to adapt to internal and external factors.
Genetic Information Flows from DNA to RNA to Protein
A gene is a stretch of DNA that serves as the blueprint for functional RNAs and proteins. Since DNA is made up of nucleotides and proteins consist of amino...
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What is Gene Expression?01:36

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A gene is a stretch of DNA that serves as the blueprint for functional RNAs and proteins. Since DNA is comprised  of nucleotides and proteins are comprised of amino acids, a mediator is required to convert the information encoded in DNA into proteins. This mediator is the messenger RNA (mRNA). mRNA copies the blueprint from DNA by a process called transcription. In eukaryotes, transcription occurs in the nucleus by complementary base-pairing with the DNA template. The mRNA is then...
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Chromatin Position Affects Gene Expression02:35

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Chromatin is the massive complex of DNA and proteins packaged inside the nucleus. The complexity of chromatin folding and how it is packaged inside the nucleus greatly influences  access to genetic information. Generally, the nucleus' periphery is considered transcriptionally repressive, while the cell's interior is considered a transcriptionally active area. 
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Gene Expression Analysis by Multiplex Single-Cell RT-PCR.

Ludovic Tricoire1, Bruno Cauli1, Bertrand Lambolez2

  • 1Sorbonne Université, UPMC Univ Paris 06 UM119, Centre National de la Recherche Scientifique (CNRS) UMR8246, Institut National de la Santé et de la Recherche Médicale (INSERM) UMRS1130, Neuroscience Paris Seine,Institut de Biologie Paris-Seine, Paris, France.

Methods in Molecular Biology (Clifton, N.J.)
|February 2, 2019
PubMed
Summary
This summary is machine-generated.

This study introduces a new protocol for analyzing gene expression in individual brain cells. This method enables simultaneous detection of 24 gene markers, offering deeper insights into cellular diversity.

Keywords:
Cell typeElectrophysiologyTranscriptomecDNAmRNA

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Area of Science:

  • Neuroscience
  • Molecular Biology
  • Genetics

Background:

  • Brain circuits are composed of diverse cellular subpopulations.
  • Understanding cellular diversity is crucial for comprehending brain function.

Purpose of the Study:

  • To develop a protocol for transcriptomic analysis of single cells post-electrophysiological recording.
  • To enable simultaneous detection of multiple gene expressions within individual neurons.

Main Methods:

  • Whole-cell patch-clamp recording and morphological reconstruction.
  • Single-cell transcriptomic analysis using multiplexed polymerase chain reaction (PCR).
  • Design and validation of gene-specific probes for PCR.

Main Results:

  • A protocol allowing transcriptomic analysis of single cells after electrophysiological recording.
  • Simultaneous detection of 24 gene/marker expressions at the single-cell level.
  • Generation of multiplexed gene expression data.

Conclusions:

  • The described protocol facilitates detailed molecular characterization of individual neurons.
  • This technique offers a powerful approach to study cellular diversity in brain circuits.
  • Provides multiplexed expression data not easily achievable with other methods.