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Related Concept Videos

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...
RACE - Rapid Amplification of cDNA Ends02:35

RACE - Rapid Amplification of cDNA Ends

Rapid Amplification of cDNA Ends, or RACE, is one of the most effective methods to obtain a full-length cDNA from an mRNA sequence between a known internal region to the unknown sequence at the 5’ or 3’ end. The unknown region is cloned in the cDNA by a gene-specific primer that binds the known end, and a hybrid primer that attaches a predefined anchor sequence to the unknown end of the cDNA. The sequence in between is amplified by PCR with an anchor primer and a gene-specific primer.
Since the...
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...
PCR01:32

PCR

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Updated: Jun 3, 2026

In Situ Hybridization for the Precise Localization of Transcripts in Plants
12:15

In Situ Hybridization for the Precise Localization of Transcripts in Plants

Published on: November 23, 2011

In situ amplification.

J O'Leary1

  • 1Department of Pathology, Cornell University Medical College, The New York Hospital-Cornell Medical Center, NY.

Methods in Molecular Medicine
|March 11, 2011
PubMed
Summary
This summary is machine-generated.

Solution phase polymerase chain reaction (PCR) cannot visualize amplified DNA in cells. In situ hybridization (ISH) allows cellular localization of nucleic acids, but conventional methods struggle with single-copy genes.

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Area of Science:

  • Molecular Biology
  • Biotechnology
  • Genetics

Background:

  • Solution phase polymerase chain reaction (PCR) is a powerful tool for amplifying DNA but lacks spatial localization capabilities within biological specimens.
  • In situ hybridization (ISH) enables the detection and localization of specific nucleic acid sequences within intact cells and tissues.
  • Conventional nonisotopic in situ detection systems often fail to detect single-copy genes, limiting their sensitivity.

Purpose of the Study:

  • To address the limitation of visualizing amplified products in cellular and tissue specimens.
  • To improve the sensitivity of in situ detection for nucleic acid sequences, including single-copy genes.

Main Methods:

  • Investigating the capabilities of in situ hybridization (ISH) for nucleic acid localization.
  • Evaluating conventional nonisotopic in situ detection systems.
  • Exploring enhanced detection techniques, such as sandwich assays, for improved sensitivity.

Main Results:

  • Solution phase PCR cannot visualize amplified products within cells or tissues.
  • In situ hybridization (ISH) permits the localization of nucleic acid sequences at the cellular level.
  • Standard nonisotopic in situ detection methods are often insufficient for detecting single-copy genes without specialized techniques.

Conclusions:

  • Visualizing amplified DNA within cells requires methods beyond solution phase PCR.
  • In situ hybridization (ISH) is a viable technique for cellular nucleic acid localization.
  • Advanced detection strategies are necessary for sensitive detection of low-abundance nucleic acid targets like single-copy genes.