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EFFECT OF DIFFERENT REGIONS OF AMPLIFIED 16S RDNA ON A PERFORMANCE OF A MULTIPLEXED, BEAD-BASED METHOD FOR ANALYSIS OF DNA SEQUENCES IN ENVIRONMENTAL SAMPLES.
Citation:
Ringbauer, J AND F J. Genthner. EFFECT OF DIFFERENT REGIONS OF AMPLIFIED 16S RDNA ON A PERFORMANCE OF A MULTIPLEXED, BEAD-BASED METHOD FOR ANALYSIS OF DNA SEQUENCES IN ENVIRONMENTAL SAMPLES. Presented at ASM Annual Meeting, Salt Lake City, UT, May 19-23, 2002.
Description:
Using a bead-based method for multiplexed analysis of community DNA, the dynamics of aquatic microbial communities can be assessed. Capture probes, specific for a genus or species of bacteria, are attached to the surface of uniquely labeled, microscopic polystyrene beads. Primers are used to amplify different regions of the 16S ribosomal DNA (rDNA) extracted from cultures or environmental samples. Specific sequences are identified by their ability to hybridize to capture probes. Hybridizations are detected by flow cytometry. To determine effect of different regions of amplified 16S rDNA, all eight variable regions in different combinations
were amplified. Capture probes, corresponding to sequences in the first third of the 16S rDNA, were used to detect various target amplicons from different populations in the environmental sample. Among the bacterial populations detected are pathogenic and fecal indicators such as Escherichia coli and Enterococcus spp. The regions amplified effected hybridization ot the capture probe, but this did not depend solely on the size of the target amplicon.