Technical Expertise
Technical expertise which would be imparted through this programme
(Hands on experience will be provided on the following)
LAB 1 : Genomics, proteomics, and bioinformatics
- DNA extraction from prokaryotes, eukaryotes and marine environmental samples
- DNA quantitation by UV Visible spectrophotometry
- Purification of DNA, agarose gel electrophoresis, Pulsed Field Gel Electrophoresis
- DNA restriction analysis
- Primer and Probe design
- PCR amplification of 16S, 18S rDNA and internal transcribed spacers, purification of the PCR products
- Ligation .
- Cloning of genes and expression studies
- Bacterial transformation, Screening of transformants by colony PCR
- Southern blotting
- Plasmid extraction, restriction digestion of the plasmid DNA
- Bacterial 16SrDNA and eukaryotic 18S rDNA and internal transcribed spacers sequencing
- DNA fingerprinting techniques - RFLP, RAPD, AFLP, microsatellite, Barcoding
- Denaturing Gradient Gel Electrophoresis
- Fluorescent In-Situ Hybridization
- RNA Isolation, cDNA synthesis, cDNA library construction and library screening and expression, Northern blotting
- Extraction and Purification of proteins
- Native & SDS PAGE, and Gradient Gel Electrophoresis,
- Western blotting, 2-Dimensional Gel electrophoresis
- Introduction to biological databases: Data mining- NCBI, EBI, DDBJ, Swiss-prot, Protein Data Bank (PDB), Protein Information Resource (PIR). Secondary databases- KEGG, Prosite, SCOP etc, and Data retrieval tools: Entrez, SRS etc, Literature search: Pubmed, Medline and OMIM etc.
- Methods in sequence analysis: pair-wise alignment -BLAST, FASTA, PSI-BLAST, flavors of BLAST (blastn, blastp,blastx, tblastx). Multiple sequence alignment-CLSUTALW, phylogenetic analysis –PHYLIP etc. Other online tools available for sequence analysis.
- Structure analysis and drug design: protein structure prediction tools-MODELLER, SWISS MODEL, and using other commercial softwares, structure visualization tools-Swiss PdbViewer, RASMOL, Cn3D etc. docking tools- ArgusLab, AutoDock and using other commercial packages. Online tools for ADME-Tox prediction
- Demonstrations
- DNA microarray technology
- DNA sequencing methods
- Lipid profiling and species identification
- Radioisotope techniques
- NMR, MS-MS, X-ray crystallography
LAB 2 : Microbiology (Bacteriology, Virology and Mycology)
- Isolation, identification and characterization (phenotypic and genotypic) of marine bacteria.
- Hydrolytic enzyme production by the marine bacterial isolates.
- 16S rRNA gene sequencing and phylogenetic analysis.
- Marine bacterial enumeration by epifluorescency.
- Isolation of marine fungi and characterization.
- Isolation of bacteriophages and plaque assay.
- Microbial community structure analysis using molecular tools.
LAB 3 : Biotechnological intervention in Aquaculture
- Disease Diagnosis and pathogen detection
- Fish sampling for laboratory observation
- Anatomy of fish and shellfish: Dissections to study the internal organs of fishes and shellfishes.
- Clinical study – skin, internal organs and gill biopsy,
- Histological methods
- Microbiological methods, Isolation of aetiological agents - identification, characterization,
- Immunodiagnostic techniques – Agglutination, Haemagglutination, Precipitation, Latex agglutination, Immuno fluorescence, Immunofluorescence histochemistry, Radial immuno diffusion,Immuno electrophoresis, Countercurrent electrophoresis and ELISA,
- DNA based diagnostic techniques: Nucleic acid probes, Dot blot hybridization, In situ hybridization, Polymerase chain reaction (PCR), RT-PCR, Nested PCR, Multiplex PCR, Broad range PCR, Real time PCR, Ribotyping, Loop Mediated isothermal Amplification (LAMP),
- Use of cell lines in disease diagnosis, Bioassay,
- Electron microscopy in disease diagnosis,
- Disease Management protocol
- Prophylaxis- Environmental management,
- Bioremediation, application of probiotics, vaccines;
- Therapy – Application of antibiotics in non-edible fish
- Water quality analysis and management
- Fish and Shell fish immunology
- Determination of total and differential haemocyte/blood count
- Prophenol oxidase activity
- NBT reduction assay
- Alkaline phosphatase and Acid phosphatase
- Lysozyme activity
- Superoxide dismutase, Catalase , Glutathione peroxidase assa
- Haemeagglutination
- Detection and quantification of antibodies in serum
- Isolation of antimicrobial peptides from shrimp haemocytes
- Cell line development and maintenance : Basic techniques in animal Cell culture: Preparation of media, Developing primary cell culture, Cell line maintenance – anchor dependent and suspension cell cultures, Subculturing, enumeration of viable cells by dye up take and exclusion, Split ratio and growth cycle, MTT assay, BrdU assay, Cytoskeleton staining, Cryo preservation,
- Application of cell lines : Virus isolation – plaque assay, Virus titration - TCID 50, Detection of viruses through immunohistochemistry/immunoflourescent staining and toxicological studies – LD 50.
- Application of Molecular markers and Marker Assisted Selection (MAS) for broodstock developments: Molecular markers – RFLP, RAPD, AFLP, Microsatellite markers, SNP, Allozymes and isozymes, Mitochondrial DNA markers, an exercise on application of DNA markers for MAS and breeding.
LAB 4 : Bioprospecting and bioprocess technology
- Collection and field identification of marine organisms
- Extraction of Bioactive compounds with different solvent systems
- Screening of extracts for bioactivity (anti-bacterial, anti viral, anti fungal and cytotoxic)
- Partitioning and enriching the bioactive compound by solid phase extraction (SPE) system
- Isolation and partial characterization of lead compound by thin layer chromatography
- Purification by column chromatography and HPLC
- Possible structure elucidation by NMR, MS-MS
- High throughput assays using cell culture: Cell viability assay, MTT assay, LDH assay
- Detection of apoptosis using DNA laddering assay and comet assay
- Screening of marine organisms for antagonistic properties using disc diffusion and microplate Quantitative assay
- Designing and optimization of culture media by Response Surface Methodology (RSM)
- Down stream processing for bioactive compounds and microbial cells
- Submerged fermentation and Solid state fermentation
- Determination of kinetics of fermentation