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Introduction to the Workshop
URLs Provided by Attendees

Abstracts

Mapping

Informatics

Sequencing

Instrumentation

Ethical, Legal, and Social Issues

Infrastructure

The electronic form of this document may be cited in the following style:
Human Genome Program, U.S. Department of Energy, DOE Human Genome Program Contractor-Grantee Workshop IV, 1994.

Abstracts scanned from text submitted for November 1994 DOE Human Genome Program Contractor-Grantee Workshop. Inaccuracies have not been corrected.

Sizing DNA Fragments by Flow Cytometry - Extending the Size Range

Jeffrey T. Petty,[1] Mitchell E. Johnson,[1] Peter M. Goodwin,[1] James H. Jett,[2] John C. Martin,[2] and Richard A. Keller[1]
[1]Chemical Science and Technology Division, MS M888, Los Alamos National Laboratory, Los Alamos, NM 87545. [2]Life Science Division, MS M888, Los Alamos National Laboratory, Los Alamos, NM 87545.

Using flow cytometry, sizes of DNA fragments were obtained from the fluorescence intensity of samples stained with a thiazole orange dye. The stained fragments passed through a low power (30 mW) continuous-wave laser beam one at a time using transit times of 1 - 5 ms. As little as 50 fg of DNA was analyzed at a rate of 40 fragments/sec in times ranging from 1 - 15 min. A detectable lower size limit of 1.5 kilobase pairs (kbp) was demonstrated, and a linear relationship between fluorescence intensity and fragment length was observed for the fragments >/= 4.4 kbp. Issues relating to size resolution in the 2 - 50 kbp range are discussed.

This work is supported by the Los Alamos Center for Human Genome Studies under United States Department of Energy Contract W-7405-ENG-36 and the National Flow Cytometry Resource, NIH RR 01315.

[1] Goodwin, P. M.; Johnson, M. E.; Martin, J. C.; Ambrose, W. P.; Marrone, B. L.; Jett, J. H.; Keller, R. A. (1993) Rapid Sizing of Individual Fluorescently Stained DNA Fragments by Flow Cytometry Nucleic Acids Res. 21, 803.
[2] Johnson, M. E.; Goodwin, P. M.; Ambrose, W. P.; Martin, J. C.; Marrone, B. L.; Jett, J. H.; Keller, R. A. (1993) Sizing of DNA Fragments by Flow Cytometry SPIE 1895, 69.