User:Dr. Poonam Sharma/Personal subpage My areas of expertise
Technical Skills Microbiology: All basic techniques, including staining, isolation, characterization and preservation of bacterial strains, chemical mutagenesis, resting cell studies. Molecular biology: Isolation and purification of chromosomal and plasmid DNA from different gram positive and gram negative bacterial strains, DNA isolation from soils and sediments, restriction digestion and cloning, DNA electrophoresis, electroporation and chemical transformation in bacteria, polymerase chain reaction (PCR), colony hybridization, southern hybridization, transposon mutagenesis. Analytical techniques: Solvent extraction of metabolites, thin layer chromatography (TLC), gas chromatography (GC), GC-MS and NMR. Protein analysis: Recombinant protein expression, analysis of protein on SDS-PAGE and protein kinetics.
Knowledge of packages used for scientific purposes like TREECON, CLUSTALX, CHEM WINDOWS for drawing chemical structures of various compounds, CLONE MANAGER for making restriction maps of DNA fragments and plasmids, and GENE RUNNER and DNASIS for analysis of DNA fragment, sequence deduction, homology searches, ORF analysis and designing of primers particularly for PCR amplification. Have experience of handling photo imaging and scanning systems and retrieval of data from instruments linked to computer. Working knowledge of ADOBE PAGE MAKER 6.5, MS-OFFICE, MS-WINDOWS (MS-WORD, MS-EXCEL, MS-POWER POINT, CORAL DRAW PHOTOSHOP etc.)
During my Ph.D. tenure I worked on various aspects of molecular microbiology. The major work was on HCH degradation where we have not only cloned genes, characterized and purified the gene products but also isolated and identified the metabolites produced by the most recalcitrant beta- and delta-HCH isomers. While doing this work, I not only learned the techniques involved in molecular biology and over expression and purification of proteins and studying the kinetics but also the various biochemistry techniques like GC, GC-MS, TLC and NMR. I also worked on another aspect “the mobilization of lin genes involved in HCH degradation pathway to a Non HCH degrader, Sphingobium chlorophenolicum DSM 7098T”. This work involves the construction of transposon cassettes, their introduction in the new host, screening of the transformants by southern hybridization and checking the activity of the introduced gene in these transformants. I am now looking forward to take up the job where I would be able to use my practical skills of molecular biology and theoretical knowledge. Research areas of my interest are Microbial genetics, physiology and metabolism, Xenobiotic degradation, Metagenomics, Microbial Ecology and Bacterial Systematic. I am sincerely interested in learning and implementing new methods, techniques and innovative ideas pertaining to microbiological research and also have a keen desire to explore the microbial diversity in terms of Structural and Functional Relationship.