Molecular Profiling of Microbial Communities from Contaminated Sources: Use of Subtractive Cloning Methods and rDNA Spacer Sequences
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.
- Research Organization:
- Univ. of Maryland, Baltimore, MD (US) (US)
- Sponsoring Organization:
- (US)
- DOE Contract Number:
- FG07-96ER62320
- OSTI ID:
- 781022
- Report Number(s):
- DOE/ER/62320; Project Number 55152; Project Number 55152; TRN: US200206%%75
- Resource Relation:
- Other Information: PBD: 10 Apr 2001
- Country of Publication:
- United States
- Language:
- English
Similar Records
Molecular Profiling of Microbial Communities from Contaminated Sources: Use of Subtractive Cloning Methods and rDNA Spacer Sequences
Molecular profiling of microbial communities from contaminated sources: Use of substractive cloning methods and rDNA spacer sequences. 1997 annual progress report
Related Subjects
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