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Rapid and Efficient FISH using Pre-Labeled Oligomer Probes.


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.

SUBMITTER: Waminal NE 

PROVIDER: S-EPMC5974128 | biostudies-literature | 2018 May

REPOSITORIES: biostudies-literature

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Rapid and Efficient FISH using Pre-Labeled Oligomer Probes.

Waminal Nomar Espinosa NE   Pellerin Remnyl Joyce RJ   Kim Nam-Soo NS   Jayakodi Murukarthick M   Park Jee Young JY   Yang Tae-Jin TJ   Kim Hyun Hee HH  

Scientific reports 20180529 1


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  ...[more]

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