Sathyamma Parvathi

• Conducted in-vitro regeneration experiments using MS medium under controlled pH, temperature, and sucrose concentrations.
• Utilized autoclave, laminar airflow cabinet, pH meter, magnetic stirrer, and incubator for sterilization, media preparation, and culture maintenance.
• Performed aseptic explant preparation using petiole base attached corm top (Dasheen & Purple Wild) and apical meristem (Mukthakeshi) under a stereo microscope.
• Studied the effects of plant growth regulators (BAP, IBA, BA, NAA) on shoot and root induction across different taro varieties.
• Identified optimal hormone concentrations for shoot induction and root induction.
• Maintained cultures in a 16/8-hour light-dark cycle under controlled incubation and transferred rooted plants to potting soil after fungicide treatment.
• Documented plant growth, hormone response, and varietal differences using digital imaging and data analysis tools.

CRISPR Protocol for loss-of-function experiment targeting gene in CASZ-1

• Conducted CRISPR/Cas9-mediated loss-of-function experiments to investigate role of CASZ1 in embryonic development, designing and synthesizing specific sgRNAs for targeted gene editing.
• Performed DNA extraction from embryonic samples, followed by PCR amplification and gel electrophoresis to confirm successful DNA editing and genotype validation.
• Utilized microinjection techniques to introduce CRISPR/Cas9 components into embryos, ensuring precise delivery and optimizing survival rates.
• Analyzed sequencing data using BioEdit and MEGA software to assess mutation efficiency, identify potential off-target effects, and validate gene modifications.
• Employed fluorescence microscopy and imaging techniques to observe phenotypic changes in embryos, documenting morphological alterations and developmental defects.

Extraction of DNA ligase from E.Coli

• Used sonication to lyse cells, applying controlled ultrasonic waves to break open the cell membranes and release the intracellular contents, ensuring efficient cell disruption.
• Followed by centrifugation to separate the lysed cell debris from the supernatant, isolating the soluble proteins, including the ligase, for further purification.
• Purified the ligase protein from the supernatant using High-Performance Liquid Chromatography (HPLC), optimizing conditions to separate ligase based on its unique properties, ensuring high purity.
• Measured the abundance of ligase through spectrophotometry, utilizing absorbance at specific wavelengths to quantify the protein concentration and assess the purity of the sample.
• Analyzed the ligase's integrity and molecular weight using SDS-PAGE gel electrophoresis, enabling the visualization of protein bands and confirmation of the expected size of the ligase protein.

Investigating Marine Bacteria for Caffeine and Xanthine Biodegradation Using 16S rRNA from Portsmouth Harbour seawater: Unveiling Caffeine Degraders and Microbial Communities

• Conducted seawater sample collection, DNA extraction, and PCR amplification for microbial analysis, optimizing bead beating and purification processes.
• Prepared samples for Illumina sequencing and processed raw data using bioinformatics tools like MOTHUR, Cutadapt, and DADA2 for taxonomic classification.
• Analyzed microbial diversity using statistical methods, including alpha and beta diversity metrics, and visualized results with MicrobiomeAnalyst.
• Identified key bacterial genera affected by caffeine and xanthine exposure, assessing their degradation potential through comparative abundance analysis.

Characterization of marine bacteria capable of NAD+ degradation via Sanger sequencing

• Cultured marine bacteria from seawater samples using the agar plate technique, isolating individual bacterial colonies for further analysis.
• Extracted DNA from bacterial isolates using standard DNA extraction methods, ensuring high-quality genomic DNA for sequencing.
• Performed gel electrophoresis to confirm the quality and integrity of the extracted DNA, ensuring successful amplification for downstream applications.
• Identified bacterial species capable of NAD+ degradation through Sanger sequencing, sequencing the DNA to obtain genetic information for further characterization.
• Analyzed Sanger sequencing data, utilizing bioinformatics tools such as Chromas and BLAST to interpret sequence results, identify gene sequences, and compare them to known databases for species identification and functional analysis.