Rapid DNA-FISH in Arabidopsis thaliana Somatic Cells

Olga Raskina, Ofir Hakim

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

Abstract

Fluorescence in situ hybridization (FISH) technique has been widely used to detect and localize specific DNA and RNA sequences in interphase nuclei and chromosomes in animals and plants. Here, we present a protocol for localization of genomic loci in nuclei of the model plant Arabidopsis thaliana. This protocol includes several advances and adaptations to A. thaliana, including preparation of nuclei and chromosomes without the use of liquid nitrogen, and an in situ hybridization procedure that preserves chromatin structure without the use of paraformaldehyde and formamide. Simultaneous denaturation of the BAC (bacterial artificial chromosome) probe and nuclei followed by annealing at high temperature allows hybridization in less than an hour. These hybridization conditions also provide high signal to noise ratio by a small number of washes. Thus, this simplified in situ hybridization procedure is completed in one working day.

Original languageAmerican English
Title of host publicationMethods in Molecular Biology
Pages259-270
Number of pages12
DOIs
StatePublished - 2024

Publication series

NameMethods in Molecular Biology
Volume2784

Keywords

  • Arabidopsis thaliana
  • BAC-FISH
  • Chromosome preparation
  • Fluorescence in situ hybridization
  • Interphase nuclei
  • Molecular cytogenetics
  • PlantPlants

All Science Journal Classification (ASJC) codes

  • Genetics
  • Molecular Biology

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