Journal
NATURE METHODS
Volume 6, Issue 2, Pages 139-141Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/NMETH.1294
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Funding
- Kirschstein-National Research Service Award
- US National Institutes of Health [GM073047, EY18082-01A2, MH080442]
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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-dimethyl-aminopropyl) 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.
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