Abstract
The utility of parallel hybridization of environmental nucleic acids to many oligonucleotides immobilized in a matrix of polyacrylamide gel pads on a glass slide (oligonucleotide microchip) was evaluated. Oligonucleotides complementary to small-subunit rRNA sequences of selected microbial groups, encompassing key genera of nitrifying bacteria, were shown to selectively retain labeled target nucleic acid derived from either DNA or RNA forms of the target sequences. The utility of varying the probe concentration to normalize hybridization signals and the use of multicolor detection for simultaneous quantitation of multiple probe-target populations were demonstrated.
Full Text
The Full Text of this article is available as a PDF (423.3 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Alm E. W., Oerther D. B., Larsen N., Stahl D. A., Raskin L. The oligonucleotide probe database. Appl Environ Microbiol. 1996 Oct;62(10):3557–3559. doi: 10.1128/aem.62.10.3557-3559.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Amann R. I., Binder B. J., Olson R. J., Chisholm S. W., Devereux R., Stahl D. A. Combination of 16S rRNA-targeted oligonucleotide probes with flow cytometry for analyzing mixed microbial populations. Appl Environ Microbiol. 1990 Jun;56(6):1919–1925. doi: 10.1128/aem.56.6.1919-1925.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Amann R. I., Ludwig W., Schleifer K. H. Phylogenetic identification and in situ detection of individual microbial cells without cultivation. Microbiol Rev. 1995 Mar;59(1):143–169. doi: 10.1128/mr.59.1.143-169.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Barns S. M., Fundyga R. E., Jeffries M. W., Pace N. R. Remarkable archaeal diversity detected in a Yellowstone National Park hot spring environment. Proc Natl Acad Sci U S A. 1994 Mar 1;91(5):1609–1613. doi: 10.1073/pnas.91.5.1609. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Boom R., Sol C. J., Salimans M. M., Jansen C. L., Wertheim-van Dillen P. M., van der Noordaa J. Rapid and simple method for purification of nucleic acids. J Clin Microbiol. 1990 Mar;28(3):495–503. doi: 10.1128/jcm.28.3.495-503.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- DeLong E. F. Archaea in coastal marine environments. Proc Natl Acad Sci U S A. 1992 Jun 15;89(12):5685–5689. doi: 10.1073/pnas.89.12.5685. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Giovannoni S. J., Britschgi T. B., Moyer C. L., Field K. G. Genetic diversity in Sargasso Sea bacterioplankton. Nature. 1990 May 3;345(6270):60–63. doi: 10.1038/345060a0. [DOI] [PubMed] [Google Scholar]
- Head I. M., Hiorns W. D., Embley T. M., McCarthy A. J., Saunders J. R. The phylogeny of autotrophic ammonia-oxidizing bacteria as determined by analysis of 16S ribosomal RNA gene sequences. J Gen Microbiol. 1993 Jun;139(Pt 6):1147–1153. doi: 10.1099/00221287-139-6-1147. [DOI] [PubMed] [Google Scholar]
- Kane M. D., Poulsen L. K., Stahl D. A. Monitoring the enrichment and isolation of sulfate-reducing bacteria by using oligonucleotide hybridization probes designed from environmentally derived 16S rRNA sequences. Appl Environ Microbiol. 1993 Mar;59(3):682–686. doi: 10.1128/aem.59.3.682-686.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Khrapko K. R., Lysov YuP, Khorlin A. A., Ivanov I. B., Yershov G. M., Vasilenko S. K., Florentiev V. L., Mirzabekov A. D. A method for DNA sequencing by hybridization with oligonucleotide matrix. DNA Seq. 1991;1(6):375–388. doi: 10.3109/10425179109020793. [DOI] [PubMed] [Google Scholar]
- Livshits M. A., Mirzabekov A. D. Theoretical analysis of the kinetics of DNA hybridization with gel-immobilized oligonucleotides. Biophys J. 1996 Nov;71(5):2795–2801. doi: 10.1016/S0006-3495(96)79473-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Maskos U., Southern E. M. Parallel analysis of oligodeoxyribonucleotide (oligonucleotide) interactions. I. Analysis of factors influencing oligonucleotide duplex formation. Nucleic Acids Res. 1992 Apr 11;20(7):1675–1678. doi: 10.1093/nar/20.7.1675. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Matulewich V. A., Strom P. F., Finstein M. S. Length of incubation for enumerating nitrifying bacteria present in various environments. Appl Microbiol. 1975 Feb;29(2):265–268. doi: 10.1128/am.29.2.265-268.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mirzabekov A. D. DNA sequencing by hybridization--a megasequencing method and a diagnostic tool? Trends Biotechnol. 1994 Jan;12(1):27–32. doi: 10.1016/0167-7799(94)90008-6. [DOI] [PubMed] [Google Scholar]
- Mobarry B. K., Wagner M., Urbain V., Rittmann B. E., Stahl D. A. Phylogenetic probes for analyzing abundance and spatial organization of nitrifying bacteria. Appl Environ Microbiol. 1996 Jun;62(6):2156–2162. doi: 10.1128/aem.62.6.2156-2162.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Poulsen L. K., Ballard G., Stahl D. A. Use of rRNA fluorescence in situ hybridization for measuring the activity of single cells in young and established biofilms. Appl Environ Microbiol. 1993 May;59(5):1354–1360. doi: 10.1128/aem.59.5.1354-1360.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Proudnikov D., Mirzabekov A. Chemical methods of DNA and RNA fluorescent labeling. Nucleic Acids Res. 1996 Nov 15;24(22):4535–4542. doi: 10.1093/nar/24.22.4535. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rees W. A., Yager T. D., Korte J., von Hippel P. H. Betaine can eliminate the base pair composition dependence of DNA melting. Biochemistry. 1993 Jan 12;32(1):137–144. doi: 10.1021/bi00052a019. [DOI] [PubMed] [Google Scholar]
- Stahl D. A., Flesher B., Mansfield H. R., Montgomery L. Use of phylogenetically based hybridization probes for studies of ruminal microbial ecology. Appl Environ Microbiol. 1988 May;54(5):1079–1084. doi: 10.1128/aem.54.5.1079-1084.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Teske A., Alm E., Regan J. M., Toze S., Rittmann B. E., Stahl D. A. Evolutionary relationships among ammonia- and nitrite-oxidizing bacteria. J Bacteriol. 1994 Nov;176(21):6623–6630. doi: 10.1128/jb.176.21.6623-6630.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Woese C. R. Bacterial evolution. Microbiol Rev. 1987 Jun;51(2):221–271. doi: 10.1128/mr.51.2.221-271.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Yershov G., Barsky V., Belgovskiy A., Kirillov E., Kreindlin E., Ivanov I., Parinov S., Guschin D., Drobishev A., Dubiley S. DNA analysis and diagnostics on oligonucleotide microchips. Proc Natl Acad Sci U S A. 1996 May 14;93(10):4913–4918. doi: 10.1073/pnas.93.10.4913. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Zheng D., Alm E. W., Stahl D. A., Raskin L. Characterization of universal small-subunit rRNA hybridization probes for quantitative molecular microbial ecology studies. Appl Environ Microbiol. 1996 Dec;62(12):4504–4513. doi: 10.1128/aem.62.12.4504-4513.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]