Document Detail


Replica plating and in situ enzymatic assay of animal cell colonies established on filter paper.
MedLine Citation:
PMID:  349559     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
We have developed a simple technique for the replica plating of Chinese hamster ovary (CHO) cells. In this procedure cells are allowed to divide for 8-16 days between the plastic surface of a petri dish and a disc of Whatman no. 50 filter paper, weighted down with glass beads. The culture medium can be replaced when necessary without disturbing the growing colonies. Cells from each developing colony grow into the fibers of the paper, while others remain attached to the plate. The cell colonies transferred to the paper are viable and can be replica plated to a new petri dish with high resolution. In this way several inositol auxotrophs have been identified in a stock of mutagen-treated cells without prior enrichment. Alternatively, the cells on the paper can be rendered permeable in situ, which permits autoradiographic screening for specific biochemical defects, as reported previously for Escherichia coli [Raetz, C. R.H. (1975 Proc. Natl. Acad. Sci. USA 72, 2274-2278]. This technique is applicable to other common cell lines and is especially useful for the identification of single colonies defective in the synthesis of DNA, RNA, protein, and membrane lipids.
Authors:
J D Esko; C R Raetz
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Proceedings of the National Academy of Sciences of the United States of America     Volume:  75     ISSN:  0027-8424     ISO Abbreviation:  Proc. Natl. Acad. Sci. U.S.A.     Publication Date:  1978 Mar 
Date Detail:
Created Date:  1978-07-15     Completed Date:  1978-07-15     Revised Date:  2009-11-18    
Medline Journal Info:
Nlm Unique ID:  7505876     Medline TA:  Proc Natl Acad Sci U S A     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  1190-3     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Cell Line
Cell Separation / methods
Clone Cells* / enzymology
Cytological Techniques*
Hela Cells / enzymology
Inositol / metabolism
L Cells (Cell Line) / enzymology
Mutation
Chemical
Reg. No./Substance:
6917-35-7/Inositol
Comments/Corrections

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


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