Automated approach for ribosomal intergenic spacer analysis of microbial diversity and its application to freshwater bacterial communities.

ABSTRACT: An automated method of ribosomal intergenic spacer analysis (ARISA) was developed for the rapid estimation of microbial diversity and community composition in freshwater environments. Following isolation of total community DNA, PCR amplification of the 16S-23S intergenic spacer region in the rRNA operon was performed with a fluorescence-labeled forward primer. ARISA-PCR fragments ranging in size from 400 to 1,200 bp were next discriminated and measured by using an automated electrophoresis system. Database information on the 16S-23S intergenic spacer was also examined, to understand the potential biases in diversity estimates provided by ARISA. In the analysis of three natural freshwater bacterial communities, ARISA was rapid and sensitive and provided highly reproducible community-specifi

SUBMITTER: Fisher MM 

PROVIDER: S-EPMC91617 | biostudies-literature | 1999 Oct

REPOSITORIES: biostudies-literature