| 摘要 |
Present invention relates to a method to determine the genotype of organisms by RAPD analysis and more specifically, to establish the relatedness of individual organisms across and within species. RAPD uses genotypic information of an organism to give an organism specific DNA fragment of different sizes. The present invention provide methods and a set of oligonucleotide primers for performing amplification and other enzymatic reactions on nucleic acid molecules that have been collected directly as environmental DNA or DNA derived form pure isolates. More specifically, the present invention relates to a novel method of genetic analysis using a set of sub-sequence, which occurs as inverted repeats in different genome with different frequencies. All bacterial cultures used in this study have been isolated from activated biomass collected from effluent treatment plants. The bacteria have been sub-cultured repeatedly to obtain pure cultures. All plating has been carried out on Luria Broth plates with 2% agar. The 16S rRNA gene has been amplified using universal primers to confirm the eubacterial nature of the isolates. The primers used to amplify a 1466-bp product were 27F forward primer 5′-AGAGTTTGATCMTGGCTCAG-3′ and 1492 reverse primer 5′-TACGGYTAC-CTTGTTACGACTT-Hence, in defined conditions two genome samples could be differentiated from each other. These features are applicable to DNA fingerprinting, marker assisted selection, genotyping, and high throughput laboratory screening methods for culturable microbes from any environmental niche.
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