Please use this identifier to cite or link to this item: https://doi.org/10.1046/j.1462-2920.2001.00233.x
Title: Optimization of an oligonucleotide microchip for microbial identification studies: A non-equilibrium dissociation approach
Authors: Liu, W.-T. 
Mirzabekov, A.D.
Stahl, D.A.
Issue Date: 2001
Source: Liu, W.-T., Mirzabekov, A.D., Stahl, D.A. (2001). Optimization of an oligonucleotide microchip for microbial identification studies: A non-equilibrium dissociation approach. Environmental Microbiology 3 (10) : 619-629. ScholarBank@NUS Repository. https://doi.org/10.1046/j.1462-2920.2001.00233.x
Abstract: The utility of a high-density oligonucleotide microarray (microchip) for identifying strains of five closely related bacilli (Bacillus anthracis, Bacillus cereus, Bacillus mycoides, Bacillus medusa and Bacillus subtilis) was demonstrated using an approach that compares the non-equilibrium dissociation rates ('melting curves') of all probe-target duplexes simultaneously. For this study, a hierarchical set of 30 oligonucleotide probes targeting the 16S ribosomal RNA of these bacilli at multiple levels of specificity (approximate taxonomic ranks of domain, kingdom, order, genus and species) was designed and immobilized in a high-density matrix of gel pads on a glass slide. Reproducible melting curves for probes with different levels of specificity were obtained using an optimized salt concentration. Clear discrimination between perfect match (PM) and mismatch (MM) duplexes was achieved. By normalizing the signals to an internal standard (a universal probe), a more than twofold discrimination (> 2.4×) was achieved between PM and 1-MM duplexes at the dissociation temperature at which 50% of the probe-target duplexes remained intact. This provided excellent differentiation among representatives of different Bacillus species, both individually and in mixtures of two or three. The overall pattern of hybridization derived from this hierarchical probe set also provided a clear 'chip fingerprint' for each of these closely related Bacillus species.
Source Title: Environmental Microbiology
URI: http://scholarbank.nus.edu.sg/handle/10635/50722
ISSN: 14622912
DOI: 10.1046/j.1462-2920.2001.00233.x
Appears in Collections:Staff Publications

Show full item record
Files in This Item:
There are no files associated with this item.

SCOPUSTM   
Citations

122
checked on Dec 14, 2017

WEB OF SCIENCETM
Citations

117
checked on Nov 18, 2017

Page view(s)

33
checked on Dec 17, 2017

Google ScholarTM

Check

Altmetric


Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.