Pulsed-field electrophoresis of megabase-sized DNA
- Department of Medicine, Stanford University Medical Center, CA (USA)
Success in constructing a physical map of the human genome will depend on two capabilities: rapid resolution of very large DNA and identification of migration anomalies. To address these issues, a systematic exploration of pulsed-field electrophoresis conditions for separating multimegabase-sized DNA was undertaken. Conditions were found for first liberating and then separating DNA up to 6 megabases at higher field strengths and more rapidly than previously reported. In addition, some conditions for transversely pulsed fields produced mobility inversion, in which increased size was accompanied by faster rather than slower migration. Importantly, anomalous migration could be identified by the presence of lateral band spreading, in which the DNA band remained sharply defined but spread laterally while moving down the gel. These results have implications for both practical applications and theoretical models of pulsed-field electrophoresis.
- OSTI ID:
- 5398279
- Journal Information:
- Molecular and Cellular Biology; (United States), Vol. 11:6; ISSN 0270-7306
- Country of Publication:
- United States
- Language:
- English
Similar Records
High-resolution separation and accurate size determination in pulsed-field gel electrophoresis of DNA. 2. Effect of pulse time and electric field strength and implications for models of the separation process
Electric birefrigence imaging of DNA in agarose electrophoresis gels
Related Subjects
DNA
ELECTROPHORESIS
GENETIC MAPPING
DNA HYBRIDIZATION
ESCHERICHIA COLI
MOLECULAR WEIGHT
PLASMIDS
SACCHAROMYCES CEREVISIAE
BACTERIA
CELL CONSTITUENTS
EUMYCOTA
FUNGI
HYBRIDIZATION
MAPPING
MICROORGANISMS
NUCLEIC ACIDS
ORGANIC COMPOUNDS
PLANTS
SACCHAROMYCES
YEASTS
550400* - Genetics