Rapid filter method for the microfluorometric analysis of DNA

✍ Scribed by Rose Ann Cattolico; Sarah P. Gibbs

• Publisher: Elsevier Science
• Year: 1975
• Tongue: English
• Weight: 656 KB
• Volume: 69
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(75)90162-1

✦ Synopsis

A rapid filter method for the microfluorometric analysis of DNA is reported in this communication. Cells collected on cellulose filters are subject to an assay sequence developed from a fluorometric method initially described by J. M. Kissane and E. Robins ((1958) J. Biol. Chem. 233, 184-188) for DNA quantitation. The assay is one of high specificity requiring no separation of DNA from other major cellular components. Samples containing as little as 0.2-0.3 pg of DNA have been analyzed by this filter method with a coefficient of variation for replicate standard samples of 3.3%. The DNA content of a number of cell types having different physiological characteristics has been determined by the use of this filter technique. The data obtained for Chlorella, Chlamydomonas, Ochromonas, and Stronglyocentrotus eggs were well within the values reported for these cells by other investigators using classical macromethods. We have taken advantage of the high specificity, sensitivity, and accuracy of this filter assay to determine DNA content during the synchronous growth cycle of the wall-less alga Olisthodiscus luteus using a single small volume culture as a sample source.