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Related Concept Videos

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...
In-situ Hybridization02:31

In-situ Hybridization

In situ hybridization (ISH) is a technique used to detect and localize specific DNA or RNA molecules in cells, tissue, or tissue sections using a labeled probe. The technique was first used in 1969 for the investigation of nucleic acids. It is currently an essential tool in scientific research and clinical settings, especially for diagnostic purposes.
Types of probes and labels
A probe is a complementary strand of DNA or RNA that binds to corresponding nucleotide sequences in a cell. Many...

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Visualization and Analysis of mRNA Molecules Using Fluorescence In Situ Hybridization in Saccharomyces cerevisiae
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Published on: June 14, 2013

Messenger RNA quantification after fluorescence-activated cell sorting using in situ hybridization.

Hiroya Yamada1, Rie Maruo, Mikio Watanabe

  • 1Department of Laboratory Medicine, Osaka University Graduate School of Medicine, Suita, Osaka 565-0871, Japan.

Cytometry. Part a : the Journal of the International Society for Analytical Cytology
|September 28, 2010
PubMed
Summary

This study introduces messenger RNA quantification after fluorescence-activated cell sorting (FACS-mQ), a new method for detecting and analyzing rare cell populations like stem cells or cancer stem cells based on gene expression.

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Method for Labeling Transcripts in Individual Escherichia coli Cells for Single-molecule Fluorescence In Situ Hybridization Experiments

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Area of Science:

  • Biotechnology
  • Molecular Biology
  • Cell Biology

Background:

  • Detecting rare cell populations (stem cells, cancer stem cells) is crucial.
  • Fluorescence-activated cell sorting (FACS) is limited by cell surface marker requirements and laborious procedures.

Purpose of the Study:

  • To establish a universal method for sorting and analyzing cells based on specific gene expression patterns.
  • To overcome the limitations of traditional FACS by enabling RNA-based cell sorting.

Main Methods:

  • Developed a protocol for messenger RNA quantification after FACS (FACS-mQ).
  • Utilized a complementary RNA (cRNA) probe for mRNA detection and staining.
  • Ensured minimal RNA degradation during the sorting process.

Main Results:

  • Successfully established the basic FACS-mQ protocol.
  • Demonstrated the potential for sorting cells based on specific mRNA expression.
  • Enabled analysis of biological characteristics via gene expression profiles.

Conclusions:

  • FACS-mQ offers a simpler and more universal approach for cell sorting and analysis.
  • This method facilitates the detection and study of stem cells and cancer stem cells in diverse tissues.
  • Overcomes FACS limitations by enabling gene expression-based cell selection.