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MiRNA in situ hybridization in formaldehyde and EDC-fixed tissues.

ABSTRACT: MicroRNAs are small regulatory RNAs with many biological functions and disease associations. We showed that in situ hybridization (ISH) using conventional formaldehyde fixation results in substantial microRNA loss from mouse tissue sections, which can be prevented by fixation with 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide that irreversibly immobilizes the microRNA at its 5' phosphate. We determined optimal hybridization parameters for 130 locked nucleic acid probes by recording nucleic acid melting temperature during ISH.

SUBMITTER: Pena JT

PROVIDER: S-EPMC2838186 | biostudies-literature | 2009 Feb

REPOSITORIES: biostudies-literature

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miRNA in situ hybridization in formaldehyde and EDC-fixed tissues.

Pena John T G JT Sohn-Lee Cherin C Rouhanifard Sara H SH Ludwig Janos J Hafner Markus M Mihailovic Aleksandra A Lim Cindy C Holoch Daniel D Berninger Philipp P Zavolan Mihaela M Tuschl Thomas T

Nature methods 20090111 2

MicroRNAs are small regulatory RNAs with many biological functions and disease associations. We showed that in situ hybridization (ISH) using conventional formaldehyde fixation results in substantial microRNA loss from mouse tissue sections, which can be prevented by fixation with 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide that irreversibly immobilizes the microRNA at its 5' phosphate. We determined optimal hybridization parameters for 130 locked nucleic acid probes by recording nucleic acid ...[more]

PMID: 19137005

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