miRNA in situ hybridization in formaldehyde and EDC–fixed tissues
JTG Pena, C Sohn-Lee, SH Rouhanifard, J Ludwig… - Nature …, 2009 - nature.com
Nature methods, 2009•nature.com
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 …
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 …
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.
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