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FISH - Fluorescent In-situ Hybridization02:07

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Fluorescence in situ hybridization, or FISH, was developed in the early 1980s and has quickly become one of the most widely used techniques in cytogenetics. Labeled probes are used to bind complementary DNA or RNA sequences on a chromosome or in a region within a cell. Earlier, the probes could only be obtained by cloning or reverse transcription of a DNA template. Currently, the probe oligonucleotides can be synthesized synthetically. Additionally, with the advancement of optical techniques,...
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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.
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A probe is a complementary strand of DNA or RNA that binds to corresponding nucleotide sequences in a cell. Many...
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DNA probes are fragments of DNA labeled with a reporter tag to enable their detection or purification. The resulting labeled DNA probes can then hybridize to target nucleic acid sequences through complementary base-pairing, and may be used to recover or identify these regions.
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Fluorescent In Situ Hybridization Using Oligonucleotide-Based Probes.

Guilherme T Braz1, Fan Yu1,2, Lívia do Vale Martins1,3

  • 1Department of Plant Biology, Michigan State University, East Lansing, MI, USA.

Methods in Molecular Biology (Clifton, N.J.)
|May 13, 2020
PubMed
Summary
This summary is machine-generated.

Oligonucleotide (oligo)-based probes offer a versatile and infinite resource for plant chromosome identification using fluorescent in situ hybridization (FISH). This method overcomes limitations of traditional probes for consistent and efficient cytogenetic studies.

Keywords:
Chromosome identificationChromosome paintingFISHOligo-FISHOligonucleotide probes

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Area of Science:

  • Plant cytogenetics
  • Molecular biology
  • Genomics

Background:

  • Chromosome identification is crucial for plant cytogenetic studies.
  • Fluorescent in situ hybridization (FISH) is a popular technique, often using bacterial artificial chromosome (BAC) clones or repetitive DNA as probes.
  • Traditional probes have limitations in identifying single chromosomes or consistency across species due to polymorphism.

Purpose of the Study:

  • To describe a protocol for oligonucleotide (oligo)-based probes for fluorescent in situ hybridization (FISH).
  • To present a versatile and efficient alternative to traditional probes for plant chromosome identification.

Main Methods:

  • Computational identification and parallel synthesis of numerous oligo probes specific to chromosomal regions or entire chromosomes.
  • Amplification and labeling of oligo-based probes.
  • Chromosome preparation and FISH procedures.

Main Results:

  • Oligo-based FISH probes are highly versatile and can identify single or multiple chromosomes.
  • Each oligo probe serves as an infinite resource for numerous FISH experiments.
  • The protocol details probe preparation, chromosome handling, and FISH execution.

Conclusions:

  • Oligo-based FISH provides a consistent and efficient method for plant chromosome identification.
  • This technique overcomes the limitations of traditional probes, offering a scalable solution for cytogenetics.
  • The described protocol facilitates the application of oligo-based FISH in research.