Single-molecule RNA fluorescence hybridization (smFISH) is a technique to visualize individual RNA molecules using multiple fluorescently-labeled oligonucleotide probes specific to the target RNA ( Raj , 2008 ; Lee , 2016a ). We adapted this technique to visualize RNAs in the whole adult worm or its germline, which enabled simultaneous recording of nascent transcripts at active transcription sites and mature mRNAs in the cytoplasm ( Lee , 2013 and 2016b). Here we describe each step of the smFISH procedure, reagents, and microscope settings optimized for extruded gonads.
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| http://dx.doi.org/10.21769/BioProtoc.2357 | DOI Listing |
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