An easy 'SteamDrop' method for high quality plant chromosome preparation
| Autor: | Mikhail G. Divashuk, Alexander Soloviev, Ilya Kirov, Katrijn Van Laere, Ludmila Khrustaleva |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2014 |
| Předmět: |
DYNAMICS
medicine.medical_specialty GENES ORGANIZATION Biology Biochemistry chemistry.chemical_compound Meiosis Genetics medicine Genetics(clinical) ALLIUM-CEPA Molecular Biology Mitosis Metaphase Genetics (clinical) New method PROTOPLASTS Biochemistry medical Steam application medicine.diagnostic_test Hybridization probe Fluorescence in situ hybridization Biochemistry (medical) Cytogenetics Methodology Biology and Life Sciences Chromosome LOCALIZATION IN-SITU HYBRIDIZATION DNA-SEQUENCES Plant chromosome preparation chemistry T-DNA Molecular Medicine DNA AIR-DRYING TECHNIQUE |
| Zdroj: | Molecular Cytogenetics MOLECULAR CYTOGENETICS |
| ISSN: | 1755-8166 |
| Popis: | Background: The chromosome preparation is a crucial step for obtaining satisfactory results in molecular cytogenetic researches. The preparation of plant chromosomes for molecular cytogenetic purposes remains a challenge for some species. In contrast to human chromosome preparation, the processes occurring during plant chromosome preparation and causing chromosome spreading are still poorly understood. Results: We studied the dynamics of plant chromosome spreading after dropping cell suspension on slides. We showed that steam stimulates cytoplasm hydrolysis and rapid chromosome spreading and that chromosomes stretch during this chromosome spreading. Based on these observations, we developed a novel method, named "SteamDrop", for the preparation of well-spread mitotic and pachytene chromosomes and successfully used it for 28 plant species with large and small chromosomes. We applied cell suspensions in ethanol instead of the commonly used ethanol/acetic acid fixative. Mitotic and meiotic chromosomes prepared via "SteamDrop" were used in fluorescent in situ hybridization (FISH) experiments with repetitive and unique DNA probes. Long storage of cell suspensions in ethanol did not impair the quality of chromosome preparations. Conclusion: The SteamDrop procedure provides a robust and routine method for high quality plant chromosome preparations. The method can be applied for metaphase as well as pachytene chromosome preparation in wide range of species. The chromosomes prepared by SteamDrop are well suitable for repetitive and unique DNA visualization. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full text from SpringerLink