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Separation of chromosomal DNA molecules from C.albicans by pulsed field gel electrophoresis.
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MedLine Citation:
PMID:  3520483     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Modifications have been made to standard pulse field gel electrophoresis (PFGE) systems to enable very large DNA molecules to be resolved. The single most important modification was to elevate the temperature of electrophoresis to 35 degrees C. This enabled the largest Saccharomyces cerevisiae chromosome to be reproducibly resolved. More impressively, it enabled the DNA of Candida albicans to be clearly resolved into six bands, a feat which was very difficult at lower temperatures. Even so, optimal resolution could only be obtained by carefully adjusting field voltages and switching times. The DNA from the two largest C. albicans chromosomes, which was estimated to be at least 5-10Mbp in size, ran somewhat anomalously, giving fuzzy bands which did not migrate in the direction of the average electric field. That the highest molecular weight band was a distinct chromosome was demonstrated by specific hybridisation to the C. albicans ADE2 gene probe. With further fine tuning, the PFGE system described here should be capable of resolving DNA from the smallest human chromosomes.
Authors:
R G Snell; R J Wilkins
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Nucleic acids research     Volume:  14     ISSN:  0305-1048     ISO Abbreviation:  Nucleic Acids Res.     Publication Date:  1986 Jun 
Date Detail:
Created Date:  1986-07-22     Completed Date:  1986-07-22     Revised Date:  2009-11-18    
Medline Journal Info:
Nlm Unique ID:  0411011     Medline TA:  Nucleic Acids Res     Country:  ENGLAND    
Other Details:
Languages:  eng     Pagination:  4401-6     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Candida albicans / analysis,  genetics*
Chromosomes / analysis*
DNA, Fungal / isolation & purification*
Electrophoresis, Agar Gel / methods
Nucleic Acid Hybridization
Saccharomyces cerevisiae / genetics
Temperature
Chemical
Reg. No./Substance:
0/DNA, Fungal
Comments/Corrections

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

Full Text
Journal Information
Journal ID (nlm-ta): Nucleic Acids Res
ISSN: 0305-1048
ISSN: 1362-4962
Article Information
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Print publication date: Day: 11 Month: 6 Year: 1986
Volume: 14 Issue: 11
First Page: 4401 Last Page: 4406
ID: 311454
PubMed Id: 3520483

Separation of chromosomal DNA molecules from C.albicans by pulsed field gel electrophoresis.
R G Snell
R J Wilkins



Article Categories:
  • Research Article


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