A general,fast, and sensitive micromethod for DNA determination: Application to rat and mouse liver,rat hepatoma,human leukocytes,chicken fibroblasts,and yeast cells |
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Authors: | Berta Fiszer-Szafarz David Szafarz Alba Guevara de Murillo |
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Institution: | Institut Curie-Biologie, Centre Universitaire, Bâtiment 111, 91405 Orsay Cédex, France |
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Abstract: | A fluorometric method using 3,5-diaminobenzoic acid for DNA determination in tissues, cultured cells, nucleated blood cells, and yeast cells is described. The method is general, simple, and rapid, and does not require prior DNA extraction, since tissue is directly solubilized in Triton X-100 and ammonia. The procedure is highly sensitive, and is able to measure rather accurately as little as 10 ng of DNA. It is applicable to all types of DNA structure. The DNA content determined in various tissues and cells was: 2.50 mg/g fresh rat liver, 3.32 mg/g rat diethylnitrosamine-induced hepatoma, 2.49 mg/g fresh mouse liver, 8.76 μg/106 human leukocytes, 3.37 μg/106 chicken fibroblasts, 2.97 μg/108 haploid yeast cells, and 2.84 μg/108 haploid yeast protoplasts. |
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