Rapid and Efficient FISH using Pre-Labeled Oligomer Probes

Scientific Reports
Nomar Espinosa WaminalHyun Hee Kim

Abstract

Fluorescence in situ hybridization (FISH) is used to visualize the distribution of DNA elements within a genome. Conventional methods for FISH take 1-2 days. Here, we developed a simplified, rapid FISH technique using pre-labeled oligonucleotide probes (PLOPs) and tested the procedure using 18 PLOPs from 45S and 5S rDNA, Arabidopsis-type telomere, and newly-identified Panax ginseng-specific tandem repeats. The 16 developed rDNA PLOPs can be universally applied to plants and animals. The telomere PLOPs can be utilized in most plants with Arabidopsis-type telomeres. The ginseng-specific PLOP can be used to distinguish P. ginseng from related Panax species. Differential labeling of PLOPs allowed us to simultaneously visualize different target loci while reducing the FISH hybridization time from ~16 h to 5 min. PLOP-FISH is efficient, reliable, and rapid, making it ideal for routine analysis, especially of newly sequenced genomes using either universal or specific targets, such as novel tandem repeats identified from whole-genome sequencing data.

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Citations

Mar 11, 2019·Chromosome Research : an International Journal on the Molecular, Supramolecular and Evolutionary Aspects of Chromosome Biology·Jiming Jiang
Jan 7, 2020·Genes & Genomics·Samantha Serafin SevillenoYoon-Jung Hwang
Jun 5, 2021·Frontiers in Plant Science·Jason SimsMarie-Therese Kurzbauer
Jul 3, 2021·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Dominika VeselinyováMiroslava Rabajdová

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Methods Mentioned

BETA
PCR

Software Mentioned

Cytovision
RepeatExplorer
Galaxy
CLC Assembly Cell
CLC Main Workbench
Adobe Photoshop CC
TAREAN

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