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Convenient and versatile DNA extraction using agarose plugs for ribotyping of problematic bacterial species.

We describe a convenient, versatile and safe method for preparing bacterial DNA for ribotyping analysis. In this method, extraction of bacterial DNA from Salmnonella typhi and Burkholderia pseudomallei. and subsequent restriction endonuclease digestion, was performed in agarose blocks/plugs thus min...

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Main Authors: Nair, S., Karim, R., Cardosa, Mary Jane, Ismail, G., Pang, T.
Format: Article
Language:English
Published: Elsevier 1999
Subjects:
Q Science (General)
QR355 Virology
R Medicine (General)
Online Access:http://ir.unimas.my/id/eprint/7114/1/Convenient.pdf
http://ir.unimas.my/id/eprint/7114/
http://www.sciencedirect.com/science/article/pii/S0167701299000755
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spelling my.unimas.ir.71142022-01-19T01:29:19Z http://ir.unimas.my/id/eprint/7114/ Convenient and versatile DNA extraction using agarose plugs for ribotyping of problematic bacterial species. Nair, S. Karim, R. Cardosa, Mary Jane Ismail, G. Pang, T. Q Science (General) QR355 Virology R Medicine (General) We describe a convenient, versatile and safe method for preparing bacterial DNA for ribotyping analysis. In this method, extraction of bacterial DNA from Salmnonella typhi and Burkholderia pseudomallei. and subsequent restriction endonuclease digestion, was performed in agarose blocks/plugs thus minimizing shearing and loss of DNA, problems commonly associated with liquid phase phenol extraction. Digested DNA in the plugs was then electrophoresed directly, transferred to nylon membranes and hybridized with labeled rDNA probes in the usual manner to provide reproducible restriction patterns. This method is particularly useful for bacterial species where standard DNA extraction in the liquid phase using phenol has been problematic (e.g. B. pseudomallei) but can be used for any bacterial species. The DNA extracted within the agarose plugs can be stored for long periods and can be used in other, widely-used typing methods such as pulsed-field gel electrophoresis (PFGE) and PCR-based techniques. Embedding live cells directly in agarose plugs also minimizes the risk of exposure to these virulent human pathogens among laboratory workers. Elsevier 1999-10 Article PeerReviewed text en http://ir.unimas.my/id/eprint/7114/1/Convenient.pdf Nair, S. and Karim, R. and Cardosa, Mary Jane and Ismail, G. and Pang, T. (1999) Convenient and versatile DNA extraction using agarose plugs for ribotyping of problematic bacterial species. Journal of Microbiological Methods, 88 (1-2). pp. 63-97. ISSN 0167-7012 http://www.sciencedirect.com/science/article/pii/S0167701299000755 10.1016/S0167-7012(99)00075-5
institution Universiti Malaysia Sarawak
building Centre for Academic Information Services (CAIS)
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Malaysia Sarawak
content_source UNIMAS Institutional Repository
url_provider http://ir.unimas.my/
language English
topic Q Science (General)
QR355 Virology
R Medicine (General)
spellingShingle Q Science (General)
QR355 Virology
R Medicine (General)
Nair, S.
Karim, R.
Cardosa, Mary Jane
Ismail, G.
Pang, T.
Convenient and versatile DNA extraction using agarose plugs for ribotyping of problematic bacterial species.
description We describe a convenient, versatile and safe method for preparing bacterial DNA for ribotyping analysis. In this method, extraction of bacterial DNA from Salmnonella typhi and Burkholderia pseudomallei. and subsequent restriction endonuclease digestion, was performed in agarose blocks/plugs thus minimizing shearing and loss of DNA, problems commonly associated with liquid phase phenol extraction. Digested DNA in the plugs was then electrophoresed directly, transferred to nylon membranes and hybridized with labeled rDNA probes in the usual manner to provide reproducible restriction patterns. This method is particularly useful for bacterial species where standard DNA extraction in the liquid phase using phenol has been problematic (e.g. B. pseudomallei) but can be used for any bacterial species. The DNA extracted within the agarose plugs can be stored for long periods and can be used in other, widely-used typing methods such as pulsed-field gel electrophoresis (PFGE) and PCR-based techniques. Embedding live cells directly in agarose plugs also minimizes the risk of exposure to these virulent human pathogens among laboratory workers.
format Article
author Nair, S.
Karim, R.
Cardosa, Mary Jane
Ismail, G.
Pang, T.
author_facet Nair, S.
Karim, R.
Cardosa, Mary Jane
Ismail, G.
Pang, T.
author_sort Nair, S.
title Convenient and versatile DNA extraction using agarose plugs for ribotyping of problematic bacterial species.
title_short Convenient and versatile DNA extraction using agarose plugs for ribotyping of problematic bacterial species.
title_full Convenient and versatile DNA extraction using agarose plugs for ribotyping of problematic bacterial species.
title_fullStr Convenient and versatile DNA extraction using agarose plugs for ribotyping of problematic bacterial species.
title_full_unstemmed Convenient and versatile DNA extraction using agarose plugs for ribotyping of problematic bacterial species.
title_sort convenient and versatile dna extraction using agarose plugs for ribotyping of problematic bacterial species.
publisher Elsevier
publishDate 1999
url http://ir.unimas.my/id/eprint/7114/1/Convenient.pdf
http://ir.unimas.my/id/eprint/7114/
http://www.sciencedirect.com/science/article/pii/S0167701299000755
_version_ 1724078454691856384
score 13.211869

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