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ArticleChromosome research : an international journal on the molecular, supramolecular and evolutionary aspects of chromosome biology2016

Fluorescent labelling of in situ hybridisation probes through the copper-catalysed azide-alkyne cycloaddition reaction.

Susann Hesse, Antonio Manetto, Valentina Cassinelli, Jörg Fuchs, Lu Ma, Nada Raddaoui, Andreas Houben

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Article in Chromosome research : an international journal on the molecular, supramolecular and evolutionary aspects of chromosome biology, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 2 papers.

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2citing papers in PubMed
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1 · What the graph read from it

What it found

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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

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3 · Its place in the literature

Who cites it

2 citing papers in PubMed.

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4 · The record

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5 · Who and what money

Authors and funding

7 authors.

Susann HesseLeibniz Institute of Plant Genetics and Crop Plant Research Gatersleben, 06466, Stadt Seeland, Germany.
Antonio Manettobaseclick GmbH, Floriansbogen 2-4, 82061, Neuried, Germany.
Valentina Cassinellibaseclick GmbH, Floriansbogen 2-4, 82061, Neuried, Germany.
Jörg FuchsLeibniz Institute of Plant Genetics and Crop Plant Research Gatersleben, 06466, Stadt Seeland, Germany.
Lu MaLeibniz Institute of Plant Genetics and Crop Plant Research Gatersleben, 06466, Stadt Seeland, Germany.
Nada RaddaouiLeibniz Institute of Plant Genetics and Crop Plant Research Gatersleben, 06466, Stadt Seeland, Germany.
Andreas HoubenLeibniz Institute of Plant Genetics and Crop Plant Research Gatersleben, 06466, Stadt Seeland, Germany. houben@ipk-gatersleben.de.

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

In situ hybridisation is a powerful tool to investigate the genome and chromosome architecture. Nick translation (NT) is widely used to label DNA probes for fluorescence in situ hybridisation (FISH). However, NT is limited to the use of long double-stranded DNA and does not allow the labelling of single-stranded and short DNA, e.g. oligonucleotides. An alternative technique is the copper(I)-catalysed azide-alkyne cycloaddition (CuAAC), at which azide and alkyne functional groups react in a multistep process catalysed by copper(I) ions to give 1,4-distributed 1,2,3-triazoles at a high yield (also called 'click reaction'). We successfully applied this technique to label short single-stranded DNA probes as well as long PCR-derived double-stranded probes and tested them by FISH on plant chromosomes and nuclei. The hybridisation efficiency of differently labelled probes was compared to those obtained by conventional labelling techniques. We show that copper(I)-catalysed azide-alkyne cycloaddition-labelled probes are reliable tools to detect different types of repetitive sequences on chromosomes opening new promising routes for the detection of single copy gene. Moreover, a combination of FISH using such probes with other techniques, e.g. immunohistochemistry (IHC) and cell proliferation assays using 5-ethynyl-deoxyuridine, is herein shown to be easily feasible.

Indexed as

AlkynesArabidopsisAzidesCell ProliferationChromosomes, PlantCopperCycloaddition ReactionDeoxyuracil NucleotidesDNA ProbesHordeumIn Situ Hybridization, FluorescenceSecaleTriticum5-ethynyl-2'-deoxyuridylic acidAlkynesAzidesCopperDeoxyuracil NucleotidesDNA ProbesChromosomesDNA sequence detectionFISHIn situ labellingInterphase

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.