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. 1969 Nov;115(3):563–568. doi: 10.1042/bj1150563

Quantitative determination of deoxyribonucleic acid in rat brain

N W Penn 1, R Suwalski 1
PMCID: PMC1185138  PMID: 5353529

Abstract

1. A procedure is given for spectrophotometric analysis of rat brain DNA after its resolution into component bases. Amounts of tissue in the range 50–100mg. can be used. 2. The amount of DNA obtained by the present method is 80% greater than that reported for rat brain by a previous procedure specific for DNA thymine. Identity of the material is established by the base ratios of purines and pyrimidines. The features responsible for the higher yield are the presence of dioxan during alkaline hydrolysis of tissue, the determination of the optimum concentration of potassium hydroxide in this step and omission of organic washes of the initial acid-precipitated residues. 3. The requirement for dioxan during alkaline hydrolysis suggests a possible association of brain DNA with lipid. The concentration of potassium hydroxide that gives maximum yield is 0·1m, indicating that there may be internucleotide linkages in this DNA that are more sensitive to alkali than those of liver or thymus DNA. 4. This procedure gives low yields of DNA from liver. It is not suitable for analysis of the DNA from this tissue.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. ALTMAN J. Autoradiographic investigation of cell proliferation in the brains of rats and cats. Anat Rec. 1963 Apr;145:573–591. doi: 10.1002/ar.1091450409. [DOI] [PubMed] [Google Scholar]
  2. ALTMAN J., DAS G. D. AUTORADIOGRAPHIC EXAMINATION OF THE EFFECTS OF ENRICHED ENVIRONMENT ON THE RATE OF GLIAL MULTIPLICATION IN THE ADULT RAT BRAIN. Nature. 1964 Dec 19;204:1161–1163. doi: 10.1038/2041161a0. [DOI] [PubMed] [Google Scholar]
  3. BURTON K. A study of the conditions and mechanism of the diphenylamine reaction for the colorimetric estimation of deoxyribonucleic acid. Biochem J. 1956 Feb;62(2):315–323. doi: 10.1042/bj0620315. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. CHARGAFF E., LIPSHITZ R., GREEN C., HODES M. E. The composition of the deoxyribonucleic acid of salmon sperm. J Biol Chem. 1951 Sep;192(1):223–230. [PubMed] [Google Scholar]
  5. DRASHER M. L. A criticism of the indiscriminate use of the Schmidt-Thannhauser method for the fractionation of nucleic acids in biological material. Science. 1953 Aug 14;118(3059):181–183. doi: 10.1126/science.118.3059.181. [DOI] [PubMed] [Google Scholar]
  6. EMANUEL C. F., CHAIKOFF I. L. Deoxyribonucleic acids of central nervous system, kidney, and spleen: a comparison of some chemical and physical properties. J Neurochem. 1960 May;5:236–244. doi: 10.1111/j.1471-4159.1960.tb13361.x. [DOI] [PubMed] [Google Scholar]
  7. HESS H. H., THALHEIMER C. MICROASSAY OF BIOCHEMICAL STRUCTURAL COMPONENTS IN NERVOUS TISSUES. I. EXTRACTION AND PARTITION OF LIPIDS AND ASSAY OF NUCLEIC ACIDS. J Neurochem. 1965 Mar;12:193–204. doi: 10.1111/j.1471-4159.1965.tb06755.x. [DOI] [PubMed] [Google Scholar]
  8. KIRBY K. S. Separation of deoxyribonucleic acid from ribonucleic acid without the use of enzymes. Biochim Biophys Acta. 1961 Feb 12;47:18–26. doi: 10.1016/0006-3002(61)90824-1. [DOI] [PubMed] [Google Scholar]
  9. KISSANE J. M., ROBINS E. The fluorometric measurement of deoxyribonucleic acid in animal tissues with special reference to the central nervous system. J Biol Chem. 1958 Jul;233(1):184–188. [PubMed] [Google Scholar]
  10. LAJTHA A. Amino acid and protein metabolism of the brain. V. Turnover of leucine in mouse tissues. J Neurochem. 1959 Feb;3(4):358–365. doi: 10.1111/j.1471-4159.1959.tb12643.x. [DOI] [PubMed] [Google Scholar]
  11. MARSHAK A. Purine and pyrimidine content of the nucleic acids of nuclei and cytoplasm. J Biol Chem. 1951 Apr;189(2):607–615. [PubMed] [Google Scholar]
  12. MESSIER B., LEBLOND C. P., SMART I. Presence of DNA synthesis and mitosis in the brain of young adult mice. Exp Cell Res. 1958 Feb;14(1):224–226. doi: 10.1016/0014-4827(58)90235-0. [DOI] [PubMed] [Google Scholar]
  13. McINDOE W. M., DAVIDSON J. N. The phosphorus compounds of the cell nucleus. Br J Cancer. 1952 Jun;6(2):200–214. [PMC free article] [PubMed] [Google Scholar]
  14. SANTEN R. J., AGRANOFF B. W. Studies on the estimation of deoxyribonucleic acid in rat brain. Biochim Biophys Acta. 1963 Jun 25;72:251–262. [PubMed] [Google Scholar]
  15. Shibko S., Koivistoinen P., Tratnyek C. A., Newhall A. R., Friedman L. A method for sequential quantitative separation and determination of protein, RNA, DNA, lipid, and glycogen from a single rat liver homogenate or from a subcellular fraction. Anal Biochem. 1967 Jun;19(3):514–528. doi: 10.1016/0003-2697(67)90242-4. [DOI] [PubMed] [Google Scholar]
  16. TAMM C., SHAPIRO H. S., LIPSHITZ R., CHARGAFF E. Distribution density of nucleotides within a desoxyribonucleic acid chain. J Biol Chem. 1953 Aug;203(2):673–688. [PubMed] [Google Scholar]
  17. TSUBOI K. K., PRICE T. D. Isolation, detection and measure of microgram quantities of labeled tissue nucleotides. Arch Biochem Biophys. 1959 Mar;81(1):223–237. doi: 10.1016/0003-9861(59)90192-4. [DOI] [PubMed] [Google Scholar]
  18. ZAMENHOF S., BURSZTYN H., RICH K., ZAMENHOF P. J. THE DETERMINATION OF DEOXYRIBONUCLEIC ACID AND OF CELL NUMBER IN BRAIN. J Neurochem. 1964 Jul;11:505–509. doi: 10.1111/j.1471-4159.1964.tb07499.x. [DOI] [PubMed] [Google Scholar]

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