I. Quantitative determination of the amount of DNA per nucleus by interference microscopy.
A method for the determination of the DNA content of isolated nuclei of different ploidy has been developed. It is based on measurement of the nuclear dry mass, with an integrating microinterferometer, before and after DNase treatment. The values found are slightly low, because, as indicated by biochemical determinations, consistently 5% to 8% of DNA is not extracted by DNase under these conditions. The average DNA values thus obtained for diploid and tetraploid nuclei of adult rat liver are 7.7 and 15.6 pg (10(-12) g), respectively. Definite advantages of this procedure are: i) comparisons with biochemical determinations to give DNA values for each class of ploidy, ii) comparisons with histophotometry of the Feulgen dye-DNA complex to give absolute values instead of arbitrary units.
D000818:Animals; D002467:Cell Nucleus / Q000032:analysis*; D003582:Cytogenetics; D004247:DNA / Q000032:analysis* / Q000378:metabolism; D004254:DNA Nucleotidyltransferases / Q000032:analysis; D004171:Diploidy; D066298:In Vitro Techniques; D008099:Liver / Q000166:cytology; D008722:Methods; D008857:Microscopy, Interference; D011123:Polyploidy
D004247:DNA; D004254:DNA Nucleotidyltransferases