Abstract
Sensitive quantification of RNA transcripts via fluorescence in situ hybridization (FISH) is a ubiquitous part of understanding quantitative gene expression in single cells. Many techniques exist to identify and localize transcripts inside the cell, but often they are costly and labor intensive. Here we present a method to use a singly labeled short DNA oligo probe to perform FISH in yeast cells. This method is effective for highly constrained FISH applications where the target length is limited (<200 nucleotides). This method can quantify different RNA isoforms or enable the use of fluorescence resonance energy transfer (FRET) to detect co-transcription of neighboring sequence blocks. Since this method relies on a single probe, it is also more cost-effective than a multiple probe labeling strategy.
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Acknowledgments
This work was supported by Georgia Institute of Technology startup funds, GAANN Molecular Biophysics and Biotechnology Fellowship, and the National Institutes of Health grant (R01-GM112882). The authors declare no conflicts of interests or competing interests.
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Wadsworth, G.M., Kim, H.D. (2022). FISHing on a Budget. In: Navid, A. (eds) Microbial Systems Biology. Methods in Molecular Biology, vol 2349. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1585-0_5
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DOI: https://doi.org/10.1007/978-1-0716-1585-0_5
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Publisher Name: Humana, New York, NY
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Online ISBN: 978-1-0716-1585-0
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