A RAPD method to detect genetic polymorphisms in fish from environmentally impacted areas
- Environmental Protection Agency, Cincinnati, OH (United States)
- Oak Ridge Inst. for Science and Education, Cincinnati, OH (United States)
- Univ. of Cincinnati, OH (United States). Dept. of Environmental Health
Random Amplified Polymorphic DNA (RAPD), an application of the polymerase chain reaction (PCR), has been used to detect genetic polymorphisms between and within species. Using the method presented here, the authors are attempting to generate statistically significant indices of population heterozygosity for Brown Bullhead catfish from two ecologically impacted inland waterways, the Black and Cuyahoga Rivers, and one reference river, Old Woman Creek. Genomic DNA, purified from nucleated red blood cells (RBCs), was randomly amplified (PCR) using 10 base oligonucleotide primers in an optimized protocol developed in this laboratory. Simplified products were electrophoretically separated in 1.65% agarose containing ethidium bromide. Following electrophoresis, agarose gels were UV irradiated and photographed with positive-negative film. The negatives were processed per manufacturer`s instructions and scanned with a densitometer. Densitometric results were then analyzed with a quantitative analysis computer program. Banding patterns among those individuals analyzed exemplify substantial polymorphisms, which may be used to assess degrees of genetic diversity in a population.
- OSTI ID:
- 40082
- Report Number(s):
- CONF-9410273-; TRN: IM9520%%277
- Resource Relation:
- Conference: 15. annual meeting of the Society of Environmental Toxicology and Chemistry (SETAC), Denver, CO (United States), 30 Oct - 3 Nov 1994; Other Information: PBD: 1994; Related Information: Is Part Of Society of Environmental Toxicology and Chemistry 15th annual meeting: Abstract book. Ecological risk: Science, policy, law, and perception; PB: 286 p.
- Country of Publication:
- United States
- Language:
- English
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