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Title: Molecular Profiling of Microbial Communities from Contaminated Sources: Use of Subtractive Cloning Methods and rDNA Spacer Sequences

Abstract

The major objective of this research was to provide appropriate sequences and assemble a DNA array of oligonucleotides to be used for rapid profiling of microbial populations from polluted areas and other areas of interest. The sequences to be assigned to the DNA array were chosen from cloned genomic DNA taken from groundwater sites having well characterized pollutant histories at Hanford Nuclear Plant and Lawrence Livermore Site 300. Glass-slide arrays were made and tested; and a new multiplexed, bead-based method was developed that uses nucleic acid hybridization on the surface of microscopic polystyrene spheres to identify specific sequences in heterogeneous mixtures of DNA sequences. The test data revealed considerable strain variation between sample sites showing a striking distribution of sequences. It also suggests that diversity varies greatly with bioremediation, and that there are many bacterial intergenic spacer region sequences that can indicate its effects. The bead method exhibited superior sequence discrimination and has features for easier and more accurate measurement.

Authors:
Publication Date:
Research Org.:
Univ. of Maryland, Baltimore, MD (US) (US)
Sponsoring Org.:
(US)
OSTI Identifier:
781022
Report Number(s):
DOE/ER/62320; Project Number 55152
Project Number 55152; TRN: US200206%%75
DOE Contract Number:  
FG07-96ER62320
Resource Type:
Technical Report
Resource Relation:
Other Information: PBD: 10 Apr 2001
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; 54 ENVIRONMENTAL SCIENCES; CLONING; COMMUNITIES; DISTRIBUTION; DNA; MIXTURES; NUCLEIC ACID HYBRIDIZATION; OLIGONUCLEOTIDES; POLLUTANTS; POLYSTYRENE; SPACERS; STRAINS; SEQUENCE; DNA ARRAYS; MICROBIAL POPULATIONS; CLONED GENOMIC DNA; ORGANIC SOLVENTS; INTERGENIC SPACER REGIONS; HYBRIDIZATION; BIOREMEDIATION

Citation Formats

Robb, Frank T. Molecular Profiling of Microbial Communities from Contaminated Sources: Use of Subtractive Cloning Methods and rDNA Spacer Sequences. United States: N. p., 2001. Web. doi:10.2172/781022.
Robb, Frank T. Molecular Profiling of Microbial Communities from Contaminated Sources: Use of Subtractive Cloning Methods and rDNA Spacer Sequences. United States. https://doi.org/10.2172/781022
Robb, Frank T. 2001. "Molecular Profiling of Microbial Communities from Contaminated Sources: Use of Subtractive Cloning Methods and rDNA Spacer Sequences". United States. https://doi.org/10.2172/781022. https://www.osti.gov/servlets/purl/781022.
@article{osti_781022,
title = {Molecular Profiling of Microbial Communities from Contaminated Sources: Use of Subtractive Cloning Methods and rDNA Spacer Sequences},
author = {Robb, Frank T},
abstractNote = {The major objective of this research was to provide appropriate sequences and assemble a DNA array of oligonucleotides to be used for rapid profiling of microbial populations from polluted areas and other areas of interest. The sequences to be assigned to the DNA array were chosen from cloned genomic DNA taken from groundwater sites having well characterized pollutant histories at Hanford Nuclear Plant and Lawrence Livermore Site 300. Glass-slide arrays were made and tested; and a new multiplexed, bead-based method was developed that uses nucleic acid hybridization on the surface of microscopic polystyrene spheres to identify specific sequences in heterogeneous mixtures of DNA sequences. The test data revealed considerable strain variation between sample sites showing a striking distribution of sequences. It also suggests that diversity varies greatly with bioremediation, and that there are many bacterial intergenic spacer region sequences that can indicate its effects. The bead method exhibited superior sequence discrimination and has features for easier and more accurate measurement.},
doi = {10.2172/781022},
url = {https://www.osti.gov/biblio/781022}, journal = {},
number = ,
volume = ,
place = {United States},
year = {Tue Apr 10 00:00:00 EDT 2001},
month = {Tue Apr 10 00:00:00 EDT 2001}
}