Ribotyping- Ribotyping of Bacterial Endophytes
Incomparison to the traditional biochemical tests, Ribotyping is a high throughput method that enables rapid identification of microorganisms. For the isolation of genomic DNA, conventional method of DNA extraction is used. The DNA thus extracted is PCR ready genomic DNA. This could be directly used for PCR without any purification step. PCR is done to amplify the gene. One set of universal primers were used for the amplification. After amplification, the PCR product was run on agarose gel. The amplicon size obtained is 1500bp. Optimization of the DNA extraction and PCR reaction was done with Escherichia coli and Bacillus subtilis (a gram –ve and gram +ve organism). In Cocosnucifera, endophytes surface sterilization was done to isolate the endophytes present in the liquid endosperm, but concluded that by doing turbidity method, it wasn’t contain any organism. Hence, concluded that coconut water is sterile. Epipremnumaureum(money plant) leaf was surface sterilized and crushed. The endophytes in leaf was isolated on nutrient agar and morphologically distinct colonies were subcultured and subjected to Ribotyping. Five colonieswere isolated from the leaf and all of them were given for sequencing to Bio-serve, Hyderabad after gel elution.