Single molecule detection: Applications to sizing of DNA fragments
- Los Alamos National Laboratory, NM (United States); and others
Using, ultrasensitive fluorescence detection and flow cytometry, size determination of ds-DNA fragments is performed using the fluorescence intensity from samples stained with a thiazole orange homodimer TOTO-1. The stained fragments pass through a low-power (30 mW) continuous-wave laser beam. Using transit times of 1-5 ms, data were acquired in times ranging from 1 to 15 mins at a rate of 40 fragments/second. As little as 50 fg of DNA was needed for the analysis. The authors have demonstrated sizing of DNA fragments in the size range from 1.5 to 150 kbp. Future applications of this approach to DNA sizing require that the factors contributing to size resolution be understood, and the authors present simulations to address this issue. To aid in the modeling, the authors have measured the saturation intensity and the relative fluorescence quantum yield of the TOTO-1/DNA complex. Applications to physical mapping of the human genome are being investigated.
- OSTI ID:
- 191743
- Report Number(s):
- CONF-941098-; TRN: 95:006733-0099
- Resource Relation:
- Conference: FACSS XXI: 21st annual conference of the Federation of Analytical Chemistry and Spectroscopy Societies (FACSS), St. Louis, MO (United States), 2-7 Oct 1994; Other Information: PBD: 1994; Related Information: Is Part Of 21st annual conference of the Federation of Analytical Chemistry and Spectroscopy Societies; PB: 257 p.
- Country of Publication:
- United States
- Language:
- English
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