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ISH: in situ hybridization protocol

Last edited Fri 12 May 2023

Stage 1 - ​Sample storage

Preserving RNA is made difficult due to presence of RNase enzyme. This enzyme is found on glassware, in reagents and on operators and their clothing. RNase quickly destroys any RNA in the cell or the RNA probe itself so users must ensure they use sterile techniques, gloves, and solutions to prevent RNase contamination of the probe or tissue RNA.

Stage 2 - Choice of probe

Stage 3 - Digoxigenin (DIG)-labeled RNA probe in situ hybridization protocol

This protocol describes the use of DIG-labeled single-stranded RNA probes to detect expression of the gene of interest in paraffin-embedded sections.