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FT
PubMed

Záznam pochází z PubMed

Plant DNA flow cytometry and estimation of nuclear genome size

Dolezel, Jaroslav
Autor Dolezel, Jaroslav Laboratory of Molecular Cytogenetics and Cytometry, Institute of Experimental Botany, Sokolovská 6, Olomouc, CZ-77200, Czech Republic. dolezel@ueb.cas.cz
Bartos, Jan
Autor Bartos, Jan

Annals of botany. 2005 Jan ; 95 (1) : 99-110.

Ann Bot
ISSN 0305-7364
Zdroj

Jazyk angličtina Země Velká Británie, Anglie Médium print

Typ dokumentu časopisecké články, práce podpořená grantem, přehledy

Perzistentní odkaz https://www.medvik.cz/link/pmid15596459

Online Plný text

PubMed 15596459
PubMed Central PMC4246710
DOI 10.1093/aob/mci005
PII: 95/1/99
Knihovny.cz E-zdroje

MeSH
buněčné jádro genetika MeSH
DNA rostlinná genetika MeSH
genetická variace MeSH
genom rostlinný * MeSH
průtoková cytometrie přístrojové vybavení metody trendy MeSH
rostliny genetika MeSH
Publikační typ
časopisecké články MeSH
práce podpořená grantem MeSH
přehledy MeSH
Názvy látek
DNA rostlinná MeSH

BACKGROUND: DNA flow cytometry describes the use of flow cytometry for estimation of DNA quantity in cell nuclei. The method involves preparation of aqueous suspensions of intact nuclei whose DNA is stained using a DNA fluorochrome. The nuclei are classified according to their relative fluorescence intensity or DNA content. Because the sample preparation and analysis is convenient and rapid, DNA flow cytometry has become a popular method for ploidy screening, detection of mixoploidy and aneuploidy, cell cycle analysis, assessment of the degree of polysomaty, determination of reproductive pathway, and estimation of absolute DNA amount or genome size. While the former applications are relatively straightforward, estimation of absolute DNA amount requires special attention to possible errors in sample preparation and analysis. SCOPE: The article reviews current procedures for estimation of absolute DNA amounts in plants using flow cytometry, with special emphasis on preparation of nuclei suspensions, stoichiometric DNA staining and the use of DNA reference standards. In addition, methodological pitfalls encountered in estimation of intraspecific variation in genome size are discussed as well as problems linked to the use of DNA flow cytometry for fieldwork. CONCLUSIONS: Reliable estimation of absolute DNA amounts in plants using flow cytometry is not a trivial task. Although several well-proven protocols are available and some factors controlling the precision and reproducibility have been identified, several problems persist: (1) the need for fresh tissues complicates the transfer of samples from field to the laboratory and/or their storage; (2) the role of cytosolic compounds interfering with quantitative DNA staining is not well understood; and (3) the use of a set of internationally agreed DNA reference standards still remains an unrealized goal.

Laboratory of Molecular Cytogenetics and Cytometry Institute of Experimental Botany Sokolovská 6 Olomouc CZ 77200 Czech Republic

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Plant DNA flow cytometry and estimation of nuclear genome size

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