High-density microarray of small-subunit ribosomal DNA probes.
Published
Journal Article
Ribosomal DNA sequence analysis, originally conceived as a way to provide a universal phylogeny for life forms, has proven useful in many areas of biological research. Some of the most promising applications of this approach are presently limited by the rate at which sequences can be analyzed. As a step toward overcoming this limitation, we have investigated the use of photolithography chip technology to perform sequence analyses on amplified small-subunit rRNA genes. The GeneChip (Affymetrix Corporation) contained 31,179 20-mer oligonucleotides that were complementary to a subalignment of sequences in the Ribosomal Database Project (RDP) (B. L. Maidak et al., Nucleic Acids Res. 29:173-174, 2001). The chip and standard Affymetrix software were able to correctly match small-subunit ribosomal DNA amplicons with the corresponding sequences in the RDP database for 15 of 17 bacterial species grown in pure culture. When bacteria collected from an air sample were tested, the method compared favorably with cloning and sequencing amplicons in determining the presence of phylogenetic groups. However, the method could not resolve the individual sequences comprising a complex mixed sample. Given these results and the potential for future enhancement of this technology, it may become widely useful.
Full Text
Duke Authors
Cited Authors
- Wilson, KH; Wilson, WJ; Radosevich, JL; DeSantis, TZ; Viswanathan, VS; Kuczmarski, TA; Andersen, GL
Published Date
- May 2002
Published In
Volume / Issue
- 68 / 5
Start / End Page
- 2535 - 2541
PubMed ID
- 11976131
Pubmed Central ID
- 11976131
International Standard Serial Number (ISSN)
- 0099-2240
Digital Object Identifier (DOI)
- 10.1128/aem.68.5.2535-2541.2002
Language
- eng
Conference Location
- United States