Meenakshi Jadhav

Project Focus :- "Site-directed mutagenesis and generation of mutant GCDH constructs of clinical mutations of Glutaric Aciduria Type I patients from Indian ethnicity."

• Performed plasmid DNA isolation of the pcDNA 6.2-GCDH wild-type construct from E. Coli transformants for downstream molecular cloning applications.
• Designed and executed oligonucleotide-based site-directed mutagenesis to generate three patient-specific mutant constructs, including both novel and known pathogenic variants in the GCDH gene.
• Gained hands-on expertise in PCR-based mutagenesis, including mutant primer design, amplification, and optimization of reaction conditions.
• Conducted molecular cloning and subcloning of wild-type GCDH from pcDNA 6.2 into the pXcmKn12 vector, strengthening skills in recombinant DNA technology and vector manipulation.
• Performed bacterial transformation and clone screening for the identification and selection of successful recombinant constructs.
• Validated cloned and mutated constructs using construct confirmation techniques, including colony PCR, restriction analysis, and Sanger sequencing.
• Interpreted Sanger sequencing chromatograms to confirm the successful incorporation of target mutations and assess sequence integrity.
• Applied in silico pathogenicity analysis using the RNAfold web server to evaluate the impact of mutations on mRNA secondary structure and stability.
• Developed a practical understanding of gene construct design, mutation validation, and genotype–phenotype correlation analysis in human disease research.
• Strengthened troubleshooting and analytical skills in experimental design, data interpretation, and documentation of molecular biology workflows.
• Gained comprehensive experience in genetic engineering and molecular diagnostics techniques, including DNA handling, cloning strategies, sequence validation, and bioinformatics-based mutation assessment.

• Gained foundational training in research methodology, literature review, and good laboratory practices (GLP), developing a strong understanding of structured scientific research and laboratory discipline.
• Acquired scientific writing skills, including the preparation of dissertations, research papers, and review articles, with practical experience contributing as a co-author on a review paper.
• Developed proficiency in Mendeley reference management software for citation formatting, and efficient organization of scientific literature.
• Performed plant and seed RNA isolation, and gained hands-on experience with essential laboratory instruments, including refrigerated centrifuges, water baths, and electronic balances.
• Trained in quantitative real-time PCR (qRT-PCR), including sample preparation, reaction setup, instrument handling, and data analysis for gene expression studies.
• Conducted agarose gel electrophoresis for quality assessment and verification of RNA and cDNA samples.
• Designed and executed a teak seed germination experiment, involving seed sowing, growth monitoring, and biological sample collection for molecular analysis.
• Carried out cDNA synthesis and differential gene expression analysis using qRT-PCR on freshly collected and pre-germinated teak seed samples.
• Performed biochemical assays to quantify antioxidant enzymes (catalase and peroxidase) and plant metabolites (total soluble proteins and sugars) using UV spectrophotometry.
• Gained introductory experience with bioinformatics tools, such as Primer3 and NCBI databases, for primer design and gene sequence analysis.
• Developed skills in experimental data interpretation and scientific report writing, integrating laboratory findings into coherent research outputs.
• Skilled at working independently and collaboratively in a team environment.

• During my internship at the ICAR-Central Agroforestry Research Institute, Jhansi, India, I had the opportunity to conduct a mini study of drought stress tolerance mechanisms of Melia dubia.
• Analyzed the morphological, physiological, and biochemical responses of 3-month-old seedlings to drought stress.
• Performed differential gene expression analysis of four known candidate genes for drought stress tolerance using real-time PCR techniques.
• Gained hands-on experience in agroforestry research, plant stress physiology, molecular biology, and data analysis.

• Completed a three-day intensive training on advanced plant biotechnology techniques.
• Gained practical experience in molecular biology and biotechnology methodologies.
• Enhanced skills in DNA isolation, PCR techniques, and plant biochemistry techniques.