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Related Concept Videos

Real Time RT-PCR02:57

Real Time RT-PCR

51.7K
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...
51.7K

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Related Experiment Video

Updated: Apr 27, 2026

Profiling Individual Human Embryonic Stem Cells by Quantitative RT-PCR
09:03

Profiling Individual Human Embryonic Stem Cells by Quantitative RT-PCR

Published on: May 29, 2014

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Profiling individual human embryonic stem cells by quantitative rt-PCR.

HoTae Lim1, In Young Choi1, Gabsang Lee2

  • 1Institute for Cell Engineering, Department of Neurology and Neuroscience, Johns Hopkins University School of Medicine.

Journal of Visualized Experiments : Jove
|June 26, 2014
PubMed
Summary
This summary is machine-generated.

Single cell qRT-PCR reveals gene expression differences in human embryonic stem cells. This method helps understand stem cell biology and population heterogeneity.

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A Combinatorial Single-cell Approach to Characterize the Molecular and Immunophenotypic Heterogeneity of Human Stem and Progenitor Populations
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Area of Science:

  • Stem cell biology
  • Molecular biology
  • Genomics

Background:

  • Stem cell population heterogeneity complicates understanding differentiation potential.
  • Single-cell transcriptome assays offer a novel approach to study individual cell variations.

Purpose of the Study:

  • To develop and validate a single-cell quantitative reverse transcription polymerase chain reaction (qRT-PCR) method.
  • To investigate gene expression heterogeneity in human embryonic stem cells at the single-cell level.

Main Methods:

  • Development of a single-cell qRT-PCR assay.
  • Sorting of OCT4::EGFP positive human embryonic stem cells.
  • Analysis of OCT4 and NANOG gene expression in individual cells.

Main Results:

  • The single-cell qRT-PCR method demonstrated effectiveness across various gene expression profiles.
  • Human embryonic stem cells showed high OCT4 expression but varied NANOG expression levels at the single-cell level.

Conclusions:

  • The developed single-cell gene expression assay is a valuable tool for analyzing stem cell population heterogeneity.
  • This method facilitates a deeper understanding of stem cell biology and differentiation potential.