Fluorescence in situ hybridization was performed with probes targeting Prrx1 (GenBank accession #NM_011127.2) mRNA by using the RNAscope Fluorescent Multiplex Detection Kit (Advanced Cell Diagnostics) according to the manufacturer instructions and as described previously (Wang et al., 2018 (link), Saraswat et al., 2021a (link)). Fixed sections were baked at 60°C for 30 minutes, washed with ethanol, tissue pretreatment with proprietary buffers. Probes were then hybridized and fluorescent conjugated. Sections were counterstained with DAPI to visualize nuclei. Positive signals were identified as punctate dots present in nucleus and cytoplasm.
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Prrx1 mRNA Detection by RNAscope
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Corresponding Organization : University at Buffalo, State University of New York
Other organizations : Aston University
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Variable analysis
independent variables
- Probes targeting Prrx1 (GenBank accession #NM_011127.2) mRNA
- Presence and location (nucleus and cytoplasm) of positive signals (punctate dots)
- Fixed sections
- Baking at 60°C for 30 minutes
- Washing with ethanol
- Tissue pretreatment with proprietary buffers
- Hybridization and fluorescent conjugation of probes
- Counterstaining with DAPI to visualize nuclei
- Not specified
- Not specified
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