Pragya Saxena
Life & science
June 2021
Microbial diversity in the natural environment is extensive. Methods for studying diversity vary and diversity can be studied at different levels. The rmophiles have been a subject of great interest as they are very important for production of industrially useful enzymes like protease, cellulose, cellulose free xylanase, amylase and also for their industrial application in dye, detergent , leather, pharmaceuticals, paper and pulp and other industries.The present study aimed for molecular identification and determination of genetic diversity of bacterial strains using random amplified polymorphic DNA (RAPD), obtained from Chhoti Anhoni hot water spring, located at pachmarhi biosphere reserve of central India . A total of 15 bacterial isolates were isolated,purified and used for molecular and genetic variability analysis. Their DNA was isolated and amplified after using two sets of universal bacterial primers after optimization. To confirm the amplification, the PCR products were loaded on an agarose gel (1.2%) and electrophoresis was carried out. Bands were visualized under UV light and documented in the gel. Amplification product of the 16s rRNA were obtained from all DNA extracted, using the PCR primers, 9F and 1542 R, which has resulted in a single band of approximately 1500bp in length and In RAPD analysis total 71 fragments were generated, of which 68 were polymorphic with an average of 6.8 bands per primer. The size of the product varied from 141bp to 2670bp. The similarity index of the isolates within each group ranged from 0.128 to 0.552. Regardless of the oligonucleotide primer employed the 15 bacterial isolates studied were separated into three genetic group composed of HSM1, HSM3, HSM4 and HSM5 (group 1), HSM 6A, HSM6B, HSM 7, HSM8, HSM9 and CA1 (group 2) and CA2, CA4, CA5, CA6 and CA7 (group 3). The PIC value ranges from 0.339 to 0.499.
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