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Direct DNA and PNA probe binding to telomeric regions without classical in situ hybridization

BACKGROUND: Fluorescence in situ Hybridization (FISH) utilizes peptide nucleic acid (PNA) probes to identify specific DNA sequences. Traditional techniques have required the heat denaturing of the DNA in formamide followed by multiple hours at moderated temperatures to allow the probe to hybridize t...

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Autores principales: Genet, Matthew D, Cartwright, Ian M, Kato, Takamitsu A
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3852893/
https://www.ncbi.nlm.nih.gov/pubmed/24103162
http://dx.doi.org/10.1186/1755-8166-6-42
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author Genet, Matthew D
Cartwright, Ian M
Kato, Takamitsu A
author_facet Genet, Matthew D
Cartwright, Ian M
Kato, Takamitsu A
author_sort Genet, Matthew D
collection PubMed
description BACKGROUND: Fluorescence in situ Hybridization (FISH) utilizes peptide nucleic acid (PNA) probes to identify specific DNA sequences. Traditional techniques have required the heat denaturing of the DNA in formamide followed by multiple hours at moderated temperatures to allow the probe to hybridize to its specific target. Over the past 30 years, advancements in both protocols and probes have made FISH a more reliable technique for both biological research and medical diagnostics, additionally the protocol has been shortened to several minutes. These PNA probes were designed to target and hybridize to both DNA and RNA, and PNA-protein interactions still remain unclear. RESULTS: In this study we have shown that a telomeric single stranded specific PNA probe is able to bind to its target without heat denaturing of the DNA and without formamide. We have also identified a centromere specific probe, which was found to bind its target with only incubation with formamide. CONCLUSIONS: Certain PNA probes are able to hybridize with their targets with minimal to no denaturing of the DNA itself. This limited denaturing preserves the chromosome structure and may lead to more effective and specific staining.
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spelling pubmed-38528932013-12-07 Direct DNA and PNA probe binding to telomeric regions without classical in situ hybridization Genet, Matthew D Cartwright, Ian M Kato, Takamitsu A Mol Cytogenet Methodology BACKGROUND: Fluorescence in situ Hybridization (FISH) utilizes peptide nucleic acid (PNA) probes to identify specific DNA sequences. Traditional techniques have required the heat denaturing of the DNA in formamide followed by multiple hours at moderated temperatures to allow the probe to hybridize to its specific target. Over the past 30 years, advancements in both protocols and probes have made FISH a more reliable technique for both biological research and medical diagnostics, additionally the protocol has been shortened to several minutes. These PNA probes were designed to target and hybridize to both DNA and RNA, and PNA-protein interactions still remain unclear. RESULTS: In this study we have shown that a telomeric single stranded specific PNA probe is able to bind to its target without heat denaturing of the DNA and without formamide. We have also identified a centromere specific probe, which was found to bind its target with only incubation with formamide. CONCLUSIONS: Certain PNA probes are able to hybridize with their targets with minimal to no denaturing of the DNA itself. This limited denaturing preserves the chromosome structure and may lead to more effective and specific staining. BioMed Central 2013-10-08 /pmc/articles/PMC3852893/ /pubmed/24103162 http://dx.doi.org/10.1186/1755-8166-6-42 Text en Copyright © 2013 Genet et al.; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Methodology
Genet, Matthew D
Cartwright, Ian M
Kato, Takamitsu A
Direct DNA and PNA probe binding to telomeric regions without classical in situ hybridization
title Direct DNA and PNA probe binding to telomeric regions without classical in situ hybridization
title_full Direct DNA and PNA probe binding to telomeric regions without classical in situ hybridization
title_fullStr Direct DNA and PNA probe binding to telomeric regions without classical in situ hybridization
title_full_unstemmed Direct DNA and PNA probe binding to telomeric regions without classical in situ hybridization
title_short Direct DNA and PNA probe binding to telomeric regions without classical in situ hybridization
title_sort direct dna and pna probe binding to telomeric regions without classical in situ hybridization
topic Methodology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3852893/
https://www.ncbi.nlm.nih.gov/pubmed/24103162
http://dx.doi.org/10.1186/1755-8166-6-42
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