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. 2024:2784:101-111.
doi: 10.1007/978-1-0716-3766-1_7.

Fluorescent In Situ Detection of Small RNAs in Plants Using sRNA-FISH

Kun Huang  1   2   3 , Blake C Meyers  4   5   6 , Jeffrey L Caplan  7   8
Affiliations

Fluorescent In Situ Detection of Small RNAs in Plants Using sRNA-FISH

Kun Huang et al. Methods Mol Biol. 2024.

Abstract

Plant small RNAs are 21-24 nucleotide, noncoding RNAs that function as regulators in plant growth and development. Colorimetric detection of plant small RNAs was made possible with the introduction of locked nucleic acid probes. However, fluorescent detection of plant small RNAs has been challenging due to the high autofluorescence from plant tissue. Here we report a fluorescent in situ detection method for plant small RNAs. This method can be applied to most plant samples and tissue types and also can be adapted for single-molecule detection of small RNAs with super-resolution microscopy.

Keywords: Autofluorescence; Confocal; FISH; In situ hybridization; Linear spectral unmixing; Plant; Quantification; RNAs; SIM; STORM; Small RNAs; Spectra.

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