[A method for determination of alkaline sucrose gradient sedimentation patterns of DNA for nondividing and slowly dividing cells].
A radiometric method for developing the alkaline sucrose gradient sedimentation patterns of DNA from non-labeled cells is described. The method is based on labeling of DNA in the gradient fractions by means of a labeled amino acid binding to DNA in the presence of formaldehyde. The procedure involves incubation of the fractions with a mixture of the labeled amino acid and formaldehyde, filtration of the incubation mixtures through nitrocellulose filters and radiometry of the filters. Relationship between the radioactivity on the filter and DNA concentration in the sample is linear. DNA detection sensitivity and specificity are satisfactory for the use under gradient ultracentrifugation condition. The non-labeled cells DNA sedimentation patterns developed by the method described and those of DNA from the same cells labeled with 3H-thimidine in vivo are found to be similar.
D000596:Amino Acids; D000818:Animals; D002286:Carcinoma, Ehrlich Tumor / Q000378:metabolism; D002417:Cattle; D002455:Cell Division; D002499:Centrifugation, Density Gradient / Q000379:methods; D055598:Chemical Phenomena; D002621:Chemistry; D004247:DNA / Q000096:biosynthesis; D004273:DNA, Neoplasm / Q000096:biosynthesis; D005557:Formaldehyde; D007553:Isotope Labeling / Q000379:methods; D051379:Mice; D013936:Thymidine / Q000378:metabolism; D013950:Thymus Gland / Q000166:cytology / Q000378:metabolism; D014316:Tritium
D000596:Amino Acids; D004273:DNA, Neoplasm; D014316:Tritium; D005557:Formaldehyde; D004247:DNA; D013936:Thymidine
Metod opredelen-ia sedimentatsionnykh raspredeleniĭ DNK v gradientakh schchelochnoĭ sakharozy, primenimyĭ k nedeliashchimsia i medlenno deliashchimsia kletkam.