DNA IMAGE-FLUOROMETRY OF INDIVIDUAL HUMAN CHROMOSOMES
N.A. Agafonova, G.A. Sakuta, Yu.M. Rozanov, G.I. Shtein, B.N. Kudryavtsev
Institute of Cytology RAS, St.-Petersburg;
e-mail: bn_kudryavtsev@mail.ru
Microfluorimetric method for the determination of DNA content in individual human chromosomes was developed. The method is based on a preliminary identification of
chromosomes with Hoechst 33258, followed by staining of the chromosomes with Feulgen reaction using Schiff's reagent type ethidium bromide-SO2, then measuring the fluorescence
intensity of the chromosomes using an image analyzer. The method allows to determine the DNA content of individual chromosomes with accuracy up to 4,5 fg. DNA content of
individual human chromosomes, their p-and q-arms as well as homologous chromosomes were measured using the developed method . It was shown that the DNA in the chromosomes
of normal human karyotype is unstable. Fluctuations of DNA in some chromosomes can reach 35-40 fg.
Key words: human chromosomes, Feulgen reaction, Schiff's reagent type ethidium bromide-SO2, DNA microfluorimetry, image analyzer
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