Document Detail


Development of a cell wash buffer that minimizes nucleic acid loss from Clostridium perfringens 10543 A.
MedLine Citation:
PMID:  2411369     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Autolytic activity and nucleic loss from Clostridium perfringens 10543 A was demonstrated during successive cell washes in hypotonic TES buffer. Autolysis increased nearly sixfold and nucleic acid loss nearly twofold when 10 mM EDTA was added to 0.3 M Tris-sucrose buffer. Attempts to minimize both autolysis and nucleic acid loss from C. perfringens during routine washing steps were unsuccessful when the effects of sucrose concentration, pH, CaCl2 addition, or wash temperature were examined independently. However, autolytic activity was eliminated and nucleic acid loss reduced to less than 5% when C. perfringens cells were washed at 4 or 25 degrees C in 1.0 M sucrose, 50 mM Tris--HCl, and 25 mM CaCl2 at pH 5.7.
Authors:
H P Blaschek; M A Klacik
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Canadian journal of microbiology     Volume:  31     ISSN:  0008-4166     ISO Abbreviation:  Can. J. Microbiol.     Publication Date:  1985 Jun 
Date Detail:
Created Date:  1985-09-30     Completed Date:  1985-09-30     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  0372707     Medline TA:  Can J Microbiol     Country:  CANADA    
Other Details:
Languages:  eng     Pagination:  575-8     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Bacteriological Techniques*
Bacteriolysis
Calcium Chloride / pharmacology
Clostridium perfringens / physiology*
DNA, Bacterial / metabolism*
Edetic Acid / pharmacology
Hydrogen-Ion Concentration
Hypotonic Solutions
Nucleic Acids / metabolism
Osmotic Pressure
RNA, Bacterial / metabolism*
Sucrose / pharmacology
Tromethamine / pharmacology
Grant Support
ID/Acronym/Agency:
1-5-24295//PHS HHS
Chemical
Reg. No./Substance:
0/DNA, Bacterial; 0/Hypotonic Solutions; 0/Nucleic Acids; 0/RNA, Bacterial; 10043-52-4/Calcium Chloride; 57-50-1/Sucrose; 60-00-4/Edetic Acid; 77-86-1/Tromethamine

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


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