AEM
Home Help [Feedback] [For Subscribers] [Archive] [Search] [Contents]
This Article
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Braun-Howland, E B
Right arrow Articles by Nierzwicki-Bauer, S A
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Braun-Howland, E B
Right arrow Articles by Nierzwicki-Bauer, S A
Agricola
Right arrow Articles by Braun-Howland, E B
Right arrow Articles by Nierzwicki-Bauer, S A

 Previous Article  |  Next Article 

Appl Environ Microbiol. 1993 October; 59(10): 3219-3224

Use of a simplified cell blot technique and 16S rRNA-directed probes for identification of common environmental isolates.

E B Braun-Howland, P A Vescio and S A Nierzwicki-Bauer

Department of Biology, Rensselaer Polytechnic Institute, Troy, New York 12180-3590.

ABSTRACT

A simple technique in which rRNA-targeted oligodeoxynucleotide probes are used to identify bacteria immobilized on membranes is described. By using colony lifts, bacteria are directly transferred from plates to untreated nitrocellulose membranes. Alternatively, cells resuspended from colonies can be applied to membranes by using a vacuum manifold under high-salt conditions. Blotted cells are baked and hybridized under stringent conditions by using standard protocols. Treatment of blotted cells with sodium dodecyl sulfate, urea, formaldehyde, or protease had no apparent effect on hybridization signals. Hybridization to rRNA from cells that had been stored refrigerated for 6 days was readily detected; however, fivefold more cells (approximately 10(7) cells) were required to obtain hybridization signals comparable to those generated by cells not subjected to storage. The sequences of oligonucleotide probes specific for Pseudomonas cepacia, Comamonas testosteroni, and Acinetobacter calcoaceticus and a group probe identifying Pseudomonas aeruginosa, Pseudomonas mendocina, Pseudomonas fluorescens, Comamonas acidovorans, and "Flavobacterium" lutescens are presented. In conjunction with the colony lift hybridization procedure, bacteria isolated from river water were identified by using these probes.

Appl Environ Microbiol. 1993 October; 59(10): 3219-3224




This article has been cited by other articles:



Home Help [Feedback] [For Subscribers] [Archive] [Search] [Contents]
J. Bacteriol. Microbiol. Mol. Biol. Rev. Eukaryot. Cell All ASM Journals

Copyright © 1993 by the American Society for Microbiology. All rights reserved.