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. 2022:2349:81-90.
doi: 10.1007/978-1-0716-1585-0_5.

FISHing on a Budget

Gable M Wadsworth  1 , Harold D Kim  2
Affiliations

FISHing on a Budget

Gable M Wadsworth et al. Methods Mol Biol. 2022.

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.

Keywords: FRET; In situ hybridization; Isoform profiling; RNA FISH; Single molecule.

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