Rapid DNA-FISH in Arabidopsis thaliana Somatic Cells.
Methods Mol Biol
; 2784: 259-270, 2024.
Article
em En
| MEDLINE
| ID: mdl-38502491
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.
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Base de dados:
MEDLINE
Assunto principal:
Arabidopsis
Limite:
Animals
Idioma:
En
Ano de publicação:
2024
Tipo de documento:
Article