miRNA in situ hybridization in mammalian tissues fixed with formaldehyde and EDC

MicroRNAs (miRNAs) are small regulatory RNAs with many biological functions and disease associations. We showed that in situ hybridization (ISH) using conventional formaldehyde fixation results in significant miRNA loss from mouse tissue sections, which can be prevented by fixation with 1–ethyl–3–(3...

Descripción completa

Detalles Bibliográficos
Autores principales: Pena, John T. G., Sohn-Lee, Cherin, Rouhanifard, Sara H., Ludwig, Janos, Hafner, Markus, Mihailovic, Aleksandra, Lim, Cindy, Holoch, Daniel, Berninger, Philipp, Zavolan, Mihaela, Tuschl, Thomas
Formato: Texto
Lenguaje:English
Publicado: 2009
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2838186/
https://www.ncbi.nlm.nih.gov/pubmed/19137005
http://dx.doi.org/10.1038/nmeth.1294
_version_ 1782178865048715264
author Pena, John T. G.
Sohn-Lee, Cherin
Rouhanifard, Sara H.
Ludwig, Janos
Hafner, Markus
Mihailovic, Aleksandra
Lim, Cindy
Holoch, Daniel
Berninger, Philipp
Zavolan, Mihaela
Tuschl, Thomas
author_facet Pena, John T. G.
Sohn-Lee, Cherin
Rouhanifard, Sara H.
Ludwig, Janos
Hafner, Markus
Mihailovic, Aleksandra
Lim, Cindy
Holoch, Daniel
Berninger, Philipp
Zavolan, Mihaela
Tuschl, Thomas
author_sort Pena, John T. G.
collection PubMed
description MicroRNAs (miRNAs) are small regulatory RNAs with many biological functions and disease associations. We showed that in situ hybridization (ISH) using conventional formaldehyde fixation results in significant miRNA loss from mouse tissue sections, which can be prevented by fixation with 1–ethyl–3–(3–dimethylaminopropyl) carbodiimide (EDC) that irreversibly immobilizes the miRNA at its 5' phosphate. We determined optimal hybridization parameters for 130 locked nucleic acid (LNA) probes by recording nucleic acid melting temperature during ISH.
format Text
id pubmed-2838186
institution National Center for Biotechnology Information
language English
publishDate 2009
record_format MEDLINE/PubMed
spelling pubmed-28381862010-03-15 miRNA in situ hybridization in mammalian tissues fixed with formaldehyde and EDC Pena, John T. G. Sohn-Lee, Cherin Rouhanifard, Sara H. Ludwig, Janos Hafner, Markus Mihailovic, Aleksandra Lim, Cindy Holoch, Daniel Berninger, Philipp Zavolan, Mihaela Tuschl, Thomas Nat Methods Article MicroRNAs (miRNAs) are small regulatory RNAs with many biological functions and disease associations. We showed that in situ hybridization (ISH) using conventional formaldehyde fixation results in significant miRNA loss from mouse tissue sections, which can be prevented by fixation with 1–ethyl–3–(3–dimethylaminopropyl) carbodiimide (EDC) that irreversibly immobilizes the miRNA at its 5' phosphate. We determined optimal hybridization parameters for 130 locked nucleic acid (LNA) probes by recording nucleic acid melting temperature during ISH. 2009-01-11 2009-02 /pmc/articles/PMC2838186/ /pubmed/19137005 http://dx.doi.org/10.1038/nmeth.1294 Text en http://www.nature.com/authors/editorial_policies/license.html#terms Users may view, print, copy, and download text and data-mine the content in such documents, for the purposes of academic research, subject always to the full Conditions of use:http://www.nature.com/authors/editorial_policies/license.html#terms
spellingShingle Article
Pena, John T. G.
Sohn-Lee, Cherin
Rouhanifard, Sara H.
Ludwig, Janos
Hafner, Markus
Mihailovic, Aleksandra
Lim, Cindy
Holoch, Daniel
Berninger, Philipp
Zavolan, Mihaela
Tuschl, Thomas
miRNA in situ hybridization in mammalian tissues fixed with formaldehyde and EDC
title miRNA in situ hybridization in mammalian tissues fixed with formaldehyde and EDC
title_full miRNA in situ hybridization in mammalian tissues fixed with formaldehyde and EDC
title_fullStr miRNA in situ hybridization in mammalian tissues fixed with formaldehyde and EDC
title_full_unstemmed miRNA in situ hybridization in mammalian tissues fixed with formaldehyde and EDC
title_short miRNA in situ hybridization in mammalian tissues fixed with formaldehyde and EDC
title_sort mirna in situ hybridization in mammalian tissues fixed with formaldehyde and edc
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2838186/
https://www.ncbi.nlm.nih.gov/pubmed/19137005
http://dx.doi.org/10.1038/nmeth.1294
work_keys_str_mv AT penajohntg mirnainsituhybridizationinmammaliantissuesfixedwithformaldehydeandedc
AT sohnleecherin mirnainsituhybridizationinmammaliantissuesfixedwithformaldehydeandedc
AT rouhanifardsarah mirnainsituhybridizationinmammaliantissuesfixedwithformaldehydeandedc
AT ludwigjanos mirnainsituhybridizationinmammaliantissuesfixedwithformaldehydeandedc
AT hafnermarkus mirnainsituhybridizationinmammaliantissuesfixedwithformaldehydeandedc
AT mihailovicaleksandra mirnainsituhybridizationinmammaliantissuesfixedwithformaldehydeandedc
AT limcindy mirnainsituhybridizationinmammaliantissuesfixedwithformaldehydeandedc
AT holochdaniel mirnainsituhybridizationinmammaliantissuesfixedwithformaldehydeandedc
AT berningerphilipp mirnainsituhybridizationinmammaliantissuesfixedwithformaldehydeandedc
AT zavolanmihaela mirnainsituhybridizationinmammaliantissuesfixedwithformaldehydeandedc
AT tuschlthomas mirnainsituhybridizationinmammaliantissuesfixedwithformaldehydeandedc

Ejemplares similares