Separation of large DNA molecules by pulsed-field capillary electrophoresis
Abstract
Pulsed-field capillary electrophoresis, using an entangled polymer solutions and permanent gels, has been employed to study the electromigration behavior of large double-stranded DNA molecules. Two different shapes of input alternating fields (sine-wave, and square-wave input signal) were examined. The electrophoretic mobilities of DNA fragments were measured as a function of the frequency of input signal and gel concentration. The resonance times were calculated from the mobility minima for each DNA chain. Differences in resonance times according to the shape of alternating electric field and employing polymer solutions or permanent gels, respectively, were observed. The separation of the 8.3-48.5 {lambda}DNA standards was obtained in approximately 45 minutes. Components of the {lambda}DNA (5-kbp ladder) were separated in less than 1 hour, while components of the {lambda}DNA (50-kbp ladder) needed approximately 3 hours for separation. The laser induced fluorescence detection of ethidium bromide stained DNA molecules was employed for the large fragments.
- Authors:
-
- Indiana Univ., Bloomington, IN (United States)
- Publication Date:
- OSTI Identifier:
- 126066
- Report Number(s):
- CONF-950402-
TRN: 95:006086-0034
- Resource Type:
- Conference
- Resource Relation:
- Conference: 209. American Chemical Society (ACS) national meeting, Anaheim, CA (United States), 2-6 Apr 1995; Other Information: PBD: 1995; Related Information: Is Part Of 209th ACS national meeting; PB: 2088 p.
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 55 BIOLOGY AND MEDICINE, BASIC STUDIES; DNA; ELECTROPHORESIS; FLUORESCENCE; GELS; MOBILITY; RESONANCE
Citation Formats
Sudor, J, and Novotny, M V. Separation of large DNA molecules by pulsed-field capillary electrophoresis. United States: N. p., 1995.
Web.
Sudor, J, & Novotny, M V. Separation of large DNA molecules by pulsed-field capillary electrophoresis. United States.
Sudor, J, and Novotny, M V. 1995.
"Separation of large DNA molecules by pulsed-field capillary electrophoresis". United States.
@article{osti_126066,
title = {Separation of large DNA molecules by pulsed-field capillary electrophoresis},
author = {Sudor, J and Novotny, M V},
abstractNote = {Pulsed-field capillary electrophoresis, using an entangled polymer solutions and permanent gels, has been employed to study the electromigration behavior of large double-stranded DNA molecules. Two different shapes of input alternating fields (sine-wave, and square-wave input signal) were examined. The electrophoretic mobilities of DNA fragments were measured as a function of the frequency of input signal and gel concentration. The resonance times were calculated from the mobility minima for each DNA chain. Differences in resonance times according to the shape of alternating electric field and employing polymer solutions or permanent gels, respectively, were observed. The separation of the 8.3-48.5 {lambda}DNA standards was obtained in approximately 45 minutes. Components of the {lambda}DNA (5-kbp ladder) were separated in less than 1 hour, while components of the {lambda}DNA (50-kbp ladder) needed approximately 3 hours for separation. The laser induced fluorescence detection of ethidium bromide stained DNA molecules was employed for the large fragments.},
doi = {},
url = {https://www.osti.gov/biblio/126066},
journal = {},
number = ,
volume = ,
place = {United States},
year = {Fri Dec 01 00:00:00 EST 1995},
month = {Fri Dec 01 00:00:00 EST 1995}
}