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Related Concept Videos

Labeling DNA Probes03:31

Labeling DNA Probes

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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.
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...
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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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DNA Staining Method Based on Formazan Precipitation Induced by Blue Light Exposure
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Fluorochromes for DNA Staining and Quantitation.

Giuliano Mazzini1,2, Marco Danova3

  • 1Institute of Molecular Genetics, CNR, Via Abbiategrasso 207, 27100, Pavia, Italy. mazzi@igm.cnr.it.

Methods in Molecular Biology (Clifton, N.J.)
|February 4, 2017
PubMed
Summary

Quantitative DNA analysis has evolved significantly, from early cytochemistry methods like the Feulgen reaction to modern flow cytometry. Advanced probes and immunofluorescence enable detailed cell analysis and clinical applications.

Keywords:
DNA contentFlow cytometryFluorescence microscopyFluorochromesQuantitative cytochemistry

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Area of Science:

  • Cell Biology
  • Molecular Biology
  • Biotechnology

Background:

  • The Feulgen reaction marked a pivotal moment in quantitative cytochemistry, initiating an era of DNA analysis.
  • Early methods relied on absorption measurements (histophotometry, cytophotometry).

Observation:

  • Fluorescence microscopy and cytofluorometry accelerated quantitative analysis of cellular components.
  • Flow cytometry initially focused on DNA quantitation, evolving with new reagents and probes.
  • Intercalating dyes and direct DNA-binding probes simplified and popularized DNA quantitation.

Findings:

  • The development of Schiff-type reagents expanded the range of fluorochromes for DNA analysis.
  • Flow cytometry advancements, including immunofluorescence, enabled multiparametric analyses.
  • Correlating DNA content with cell markers provides insights into cell cycle and subpopulations.

Implications:

  • Flow cytometry has become crucial in clinical applications for analyzing cell subpopulations.
  • Multiparametric analyses in flow cytometry aid in defining cytokinetic characteristics based on immunophenotype.
  • The evolution of DNA quantitative analysis showcases advancements in cell biology research and diagnostics.