Suzuki, M. T.. Monterey Bay Aquarium Research Institute, suzuki@mbari.org
Taylor, L. T.. Monterey Bay Aquarium Research Institute, taylor@mbari.org
DeLong, E. F.. Monterey Bay Aquarium Research Institute, delong@mbari.org

 
QUANTITATIVE ANALYSIS OF SMALL SUBUNIT RIBOSOMAL RNA GENES IN MIXED MICROBIAL POPULATIONS USING 5' NUCLEASE ASSAYS.
 
Ribosomal RNA genes (rDNAs) are extensively used to study the diversity of microorganisms in nature. Several methods are currently used to estimate the frequency of specific rDNAs in environmental samples. However, these methods are labor intensive, require large samples and yield measurements that are semi-quantitative at best. In this study we applied 5' nuclease assays to estimate relative proportions of different 16S rDNAs in picoplankton samples from the Monterey Bay. We compared the proportion of 16S rDNAs from Bacteria, and Archaea estimated by 5' nuclease assays, to those estimated by oligonucleotide hybridization to rRNA immobilized on membranes, and by in situ hybridization. We also compared the relative proportion of 16S rDNAs of oxygenic phototrophic bacteria estimated by 5' nuclease assays (Synechococcus and Prochlorococcus) to the relative cell numbers estimated by flow cytometry. The results show good agreement between relative proportions of 16S rDNAs estimated by 5' nuclease assays compared to oligonucleotide hybridization or cell counts. The data indicate that 5' nuclease assays will be useful for rapid, high throughput quantification of specific rDNAs in environmental samples.
 
Day: Tuesday, Feb. 2
Time: Poster
Location: Sweeney Center
 
Code: SS40TU1074S