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Related Concept Videos

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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Observation and Quantification of Telomere and Repetitive Sequences Using Fluorescence In Situ Hybridization FISH with PNA Probes in Caenorhabditis elegans
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Telomere Aberration Detection by PNA FISH Probe.

Takamitsu A Kato1

  • 1Department of Environmental & Radiological Health Sciences, Colorado State University, Fort Collins, CO, USA. Takamitsu.Kato@colostate.edu.

Methods in Molecular Biology (Clifton, N.J.)
|September 6, 2022
PubMed
Summary

Telomeres shorten with cell division and aging, potentially causing genomic instability. This study details a PNA FISH protocol to visualize telomere signals on metaphase chromosomes, aiding in the study of telomere dysfunction.

Keywords:
End cap of chromosomesHeyflick’s limitPNA FISHTelomereTelomere fusionTelomeric repeat sequences

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Area of Science:

  • Genetics
  • Cell Biology
  • Molecular Biology

Background:

  • Telomeres are protective caps at chromosome ends.
  • Telomere shortening occurs with cell division and aging.
  • Shortened telomeres can lead to fusion and genomic instability.

Purpose of the Study:

  • To introduce a PNA FISH protocol for telomere visualization.
  • To provide a method for observing telomere signals on metaphase chromosomes.

Main Methods:

  • Utilizing PNA (peptide nucleic acid) FISH probes targeting telomere repeat sequences (TTAGGG)n.
  • Adapting traditional or modified FISH protocols for telomere staining.
  • Applying the protocol to metaphase chromosomes.

Main Results:

  • The PNA FISH protocol effectively visualizes telomere signals.
  • PNA probes demonstrate higher hybridization affinity compared to DNA probes.
  • This method offers improved telomere staining on metaphase chromosomes.

Conclusions:

  • The PNA telomere FISH protocol is a valuable tool for studying telomere length and dysfunction.
  • This technique facilitates the assessment of genomic instability related to telomere shortening.