Advanced Training Module

Microbiology Techniques – Advance

Module 1

Duration: 15 Days

Training fees: `3500/-

Course Content

• Basics of Lab Safety: Lab safety and Procedures, Scope & Applications, Record Maintenance, Handling of Equipment

• Overview and types of Sterilization techniques, Concept of Solution Chemistry and Concentration, Keeping & storage of lab chemicals, reagents & solvents

• Type of Media, Media Preparation (NAM, EMB, MSA, etc.,)

• Sample collection and Serial Dilution technique

• Culture techniques (pour plate, spread plate and streak plates methods)

• Pure culture preparation (bacteria and fungi)

• Gram’s staining, Endospore staining, Lactophenol aniline cotton blue staining.

• Microscopy, correct methods of handling microscope.

• Characterization of Bacteria by Biochemical Tests;

1. 1. Catalase test
   2. Indole test
   3. Methyl red reduction test,
   4. Voges Proskauer test
   5. Citrate utilization test

Microbiology Techniques – Advance

Module 2

Duration: 30 Days

Training fees: ` 8000/-

Course Content

• Basics of Lab Safety: Lab safety and Procedures, Scope & Applications, Record Maintenance, Handling of Equipment

• Overview and types of Sterilization techniques, Concept of Solution Chemistry and Concentration, Keeping & storage of lab chemicals, reagents & solvents

• Type of Media, Media Preparation (NAM, EMB, MSA, XLD, BHI etc.,)

• Sample collection and Serial Dilution technique

• Culture techniques (pour plate, spread plate and streak plates methods)

• Pure culture preparation (bacteria and fungi)

• Gram’s staining, Endospore staining, Lactophenol aniline cotton blue staining.

• Microscopy, correct methods of handling microscope.

• Characterization of Bacteria by Biochemical Tests;

1. 1. 1. Catalase Test
      2. Bacterial Motility Test
      3. IMViC Test
      4. Starch Hydrolysis Test
      5. Gelatin Hydrolysis Test
      6. Urease Test
      7. Carbohydrate Fermentation Test (glucose, sucrose, lactose, mannitol, dextrose etc.)

• Antimicrobial Activity;

1. 1. Preparation of NAM/Muller Hinton agar media
   2. Antimicrobial susceptibility assay by well diffusion & disc diffusion methods
   3. Antimicrobial sensitivity assay (MIC assay) by well diffusion methods.

Biochemistry & Bio-Analytical Techniques – Advance

Module 3

Duration: 15-20 Days

Training fees: ` 10000/-

Course Content

• Concept of Biochemistry, Scope & Applications.

• Basics of Lab Safety: Lab safety and Procedures, Record Maintenance, Handling of Equipment

• Quantitative estimation of Proteins by Lowry’s method (Spectrophotometric)

• Estimation of Total carbohydrates by Anthron or DNS method (Spectrophotometric)

• Estimation of Total fats

• Estimation of Saponification value of Fats or Oils

• Estimation of Ascorbic acid in food or beverage samples

• Thin Layer Chromatography analysis of extracts for qualitative detection of marker compounds (phenolic, flavonoid or amino acid).

• Column Chromatography technique for separation of mixture of chemical constituents

• HPLC Analysis for detection of marker;

1. 1. HPLC Machine overview
   2. Scope of HPLC analysis in food, pharma, medical, environment and agriculture sector
   3. Sample preparation for HPLC analysis
   4. Preparation of standard plot of standard compound/marker
   5. Detection and quantification of marker compound in test samples

Molecular Biology Techniques – Advance

Module 4

Duration: 15 Days

Training fees: ` 10000/-

Course Content

• Basics of Lab Safety: Lab safety and Procedures, Scope & Applications, Record Maintenance, Handling of Equipment, 

• Sterilization Techniques, Preparation of Chemical & Reagents.

• Extraction of DNA from Plant samples by CTAB method

• Extraction of Bacterial DNA by PCI method

• Extraction of Human DNA from Blood samples

• Qualitative and quantitative analysis of extracted DNA by Electrophoresis on 2% agarose gel and by Spectrophotometry.

• PCR Techniques and protocol for extracted DNA samples

• RAPD Fingerprinting Techniques using PCR

• Agarose gel electrophoresis & gel documentation for detection amplified DNA fragment.

Molecular Biology Techniques – Advance

Module 5

Duration: 30 Days

Training fees: `15000/-

Course Content

• Basics of Lab Safety: Lab safety and Procedures, Scope & Applications, Record Maintenance, Handling of Equipment, 

• Sterilization Techniques, Preparation of Chemical & Reagents.

• Extraction of DNA from Plant samples by CTAB method

• Extraction of Bacterial DNA by PCI method

• Extraction of Human DNA from Blood samples

• Metagenomic DNA extraction 

• Qualitative and quantitative analysis of extracted DNA by Electrophoresis on 2% agarose gel and by Spectrophotometry.

• PCR Primer designing, In-Silico PCR and Practical use of Bioinformatics

• PCR Techniques and protocol for extracted DNA samples

• Agarose gel electrophoresis & gel documentation for detection amplified DNA fragment.

• RFLP and RAPD Fingerprinting Techniques using PCR

• 16S-rRNA amplification and BLAST analysis of 16S-rRNA sequence for bacterial identification.

Recombinant DNA Technology & Genetic Engineering

Module 6

Duration: 15 to 20 Days

Training fees: `10000/-

Course Content

• Basics of Lab Safety: Lab safety and Procedures, Scope & Applications, Record Maintenance, Handling of Equipment 

• Sterilization Techniques, Preparation of Chemical & Reagents.

• Preparation of LB media and culture of Culture of E.coli bacteria

• Extraction of Bacterial Genomic DNA

• Extraction of Plasmid DNA from Bacterial cells

• Qualitative and quantitative analysis of extracted DNA by Electrophoresis on 2% agarose gel and by Spectrophotometry.

• Preparation of Competent cells

• Restriction digestion of λ DNA using restriction endonucleases Hind III/ EcoR I

• Used of T4 DNA ligase and construction of vector DNA.

• Agarose gel electrophoresis for confirmation of cutting and ligation of sample DNA

Recombinant DNA Technology & Genetic Engineering

Module 7

Duration: 30 Days

Training fees: `25000/-

Course Content

• Basics of Lab Safety: Lab safety and Procedures, Scope & Applications, Record Maintenance, Handling of Equipment 

• Sterilization Techniques, Preparation of Chemical & Reagents.

• Preparation of LB media and culture of Culture of E.coli bacteria

• Extraction of Bacterial Genomic DNA

• Extraction of Plasmid DNA from Bacterial cells

• Qualitative and quantitative analysis of extracted DNA by Electrophoresis on 2% agarose gel and by Spectrophotometry.

• Preparation of Competent cells

• Restriction digestion of λ DNA using restriction endonucleases Hind III/ EcoR I

• Used of T4 DNA ligase and construction of vector DNA 

• Cloning Transformation and Blue White Selection

• Colony PCR to Amplify λ DNA

• IPTG Induced Gene expression in Bacterial cells by SDS-PAGE Analysis

Medical Biotech/Microbiology: Antibiotic Sensitivity Assay Methods

Basic Module 8

Duration: 15 – 20 Days

Training fees: ` 5000/-

Course Content

• Basics of Lab Safety: Lab safety and Procedures, Scope & Applications, Record Maintenance, Handling of Equipment

• Overview and types of Sterilization techniques, Concept of Solution Chemistry and Concentration, Keeping & storage of lab chemicals, reagents & solvents

• Concept of Antimicrobial Agents: Types of antimicrobial compounds, Antibiotics vs. antifungals vs. herbal extracts and Bactericidal vs. bacteriostatic action

• Media & Reagent Preparation for AST

1. 1. 1. Pre-experimental preparations
      2. Mueller Hinton Agar (MHA) preparation
      3. Nutrient Broth, Nutrient Agar
      4. Sterilization and storage of media, discs, and filters

• Preparation of Test Microorganisms

1. 1. 1. Revival and maintenance of bacterial cultures
      2. Preparation of inoculum (0.5 McFarland Standard)
      3. Spread plating for lawn culture

• Antibiotic Sensitivity Testing Methods: Kirby–Bauer Disc Diffusion Technique, Agar Well Diffusion Method, Determination of Zone of Inhibition

• Minimum Inhibitory Concentration (MIC) – Basics: Broth dilution overview, Serial dilution of antimicrobial agents/extracts, Observation of turbidity and interpretation

• Hands-on with Antibiotic Discs & Herbal Extracts

1. 1. 1. Placing and labelling antibiotic discs
      2. Loading wells with extracts
      3. Incubation, observation, zone diameter measurement

• Clinical Sample Testing

1. 1. 1. Overview of blood, urine, pus, wound samples cultures
      2. Practical handling and antibiotic sensitivity assay with standard antibiotics

• Result Interpretation

1. 1. Measuring zones and reporting using standard tables
   2. CLSI standard interpretation (Sensitive / Intermediate / Resistant)
   3. Report writing and data representation (tables, graphs)

Medical Biotechnology: Molecular Diagnostics: Gene-Specific Detection of Pathogens

Module 9

Duration: 10-15 Days

Training fees: `10000/-

Course Content

1. Concept of Molecular Diagnostics; gene markers for pathogen identification (16S-rRNA, ITS region, virulence and coat protein genes), scope & applications in clinical, medical biotechnology.
2. Basics of Lab Safety: Biosafety levels, aseptic handling, record maintenance, contamination control, handling of instruments.
3. Sample Collection and Processing: Soil/water samples, microbial cultures; transport media and storage conditions.
4. Extraction of pathogen genomic DNA (bacterial/fungal) by PCI method.
5. Extraction of viral RNA by TRIzol/phenol method.
6. Qualitative and quantitative analysis of extracted DNA/RNA by agarose gel electrophoresis and spectrophotometry.
7. Primer selection and PCR reaction setup for gene-specific detection; master mix preparation, template addition, positive & negative controls.
8. Gradient PCR for optimization of annealing temperature of primers.
9. Agarose gel electrophoresis & gel documentation for visualization of amplified gene fragment.
10. Interpretation of PCR results; calculation of amplicon size, troubleshooting false positives/negatives.
11. Report preparation for gene-specific pathogen detection.
12. Clinical/Industrial documentation: SOP writing, batch record, QC/QA basics.

Medical Biotechnology: Molecular Diagnostics: Gene-Specific Detection of Pathogens

Module 10

Duration: 30 Days

Training fees: `20000/-

Course Content

1. Concept of Molecular Diagnostics; gene markers for pathogen identification (16S-rRNA, ITS region, virulence and coat protein genes), scope & applications in clinical, medical biotechnology.
2. Basics of Lab Safety: Biosafety levels, aseptic handling, record maintenance, contamination control, handling of instruments.
3. Sample Collection and Processing: Soil/water samples, microbial cultures; transport media and storage conditions.
4. Extraction of pathogen genomic DNA (bacterial/fungal) by PCI method.
5. Extraction of viral RNA by TRIzol/phenol method.
6. Qualitative and quantitative analysis of extracted DNA/RNA by agarose gel electrophoresis and spectrophotometry.
7. Gene-Specific Primer Design: for target pathogen gene

• Identifying unique genes (toxins, virulence, coat proteins, housekeeping)
• NCBI gene search, FASTA download
• Primer design on Primer-BLAST
• In-silico PCR & amplicon size prediction

1. PCR reaction setup for gene-specific detection; master mix preparation, template addition, positive & negative controls.
2. Gradient PCR for optimization of annealing temperature of primers.
3. Agarose gel electrophoresis & gel documentation for visualization of amplified gene fragment.
4. Advanced Detection Methods (demonstration or hands-on)

1. RT-PCR for RNA viruses 
2. Multiplex PCR for detection of multiple pathogens 
3. RAPD/RFLP demonstration for differentiation 

1. Interpretation of PCR results; calculation of amplicon size, troubleshooting false positives/negatives.
2. Report preparation for gene-specific pathogen detection.
3. Clinical/Industrial documentation: SOP writing, batch record, QC/QA basics.

Pharma Biotechnology: Phyto Drug Extraction Technology

Module 11

Duration: 15 Days

Training fees: ` 6000/-

Course Content

• Concept of Phytodrug and Pharmaceutical Biotechnology, Scope & Applications.

• Basics of Lab Safety: Lab safety and Procedures, Record Maintenance, Handling of Equipment

• Collection of plant material for drug extraction

• Extraction of phytochemicals by maceration, infusion and Soxhletion methods

• Concentration of Phytochemical Extracts and organoleptic assessment

• Preliminary qualitative test to detect different bioactive chemical constituents;

1. 1. 1. Alkaloid test
      2. Flavonoid test
      3. Tannin test
      4. Terpenoid test
      5. Glycoside test
      6. Saponin test
      7. Anthraquinone test

• Thin Layer Chromatography analysis of extracts for qualitative detection of marker compounds (quercetin or gallic acid).

• Quantitative estimation of bioactive constituents by Spectrophotometric methods;

1. 1. Estimation of total polyphenolics (gallic acid equivalent) in phytoextract
   2. Estimation of total flavonoids (quercetin equivalent) in phytoextract

Pharma Biotechnology: Phyto Drug Extraction Technology & Analysis

Module 12

Duration: 15 Days

Training fees: ` 10000/-

Course Content

• Concept of Phytodrug and Pharmaceutical Biotechnology, Scope & Applications.

• Basics of Lab Safety: Lab safety and Procedures, Record Maintenance, Handling of Equipment

• Collection of plant material for drug extraction

• Extraction of phytochemicals by maceration, infusion and Soxhletion methods

• Concentration of Phytochemical Extracts and organoleptic assessment

• Preliminary qualitative test to detect different bioactive chemical constituents;

1. 1. 1. Alkaloid test
      2. Flavonoid test
      3. Tannin test
      4. Terpenoid test
      5. Glycoside test
      6. Saponin test
      7. Anthraquinone test

• Thin Layer Chromatography analysis of extracts for qualitative detection of marker compounds (quercetin or gallic acid).

• Quantitative estimation of bioactive constituents by Spectrophotometric methods;

1. 1. 1. Estimation of total polyphenolics (gallic acid equivalent) in phytoextract
      2. Estimation of total flavonoids (quercetin equivalent) in phytoextract

• HPLC Analysis of phytochemical extracts;

1. 1. HPLC Machine overview
   2. Sample preparation for HPLC analysis
   3. Preparation of standard plot of standard compound/marker
   4. Detection and quantification of marker compound in test samples

Pharma Biotechnology: Phyto Drug Extraction, Analysis & Biological Activity

Module 13

Duration: 20-25 Days

Training fees: ` 15000/-

Course Content

• Concept of Phytodrug and Pharmaceutical Biotechnology, Scope & Applications.

• Basics of Lab Safety: Lab safety and Procedures, Record Maintenance, Handling of Equipment

• Collection of plant material for drug extraction

• Extraction of phytochemicals by maceration, infusion and Soxhletion methods

• Concentration of Phytochemical Extracts and organoleptic assessment

• Preliminary qualitative test to detect different bioactive chemical constituents;

1. 1. 1. Alkaloid test
      2. Flavonoid test
      3. Tannin test
      4. Terpenoid test
      5. Glycoside test
      6. Saponin test

• Thin Layer Chromatography analysis of extracts for qualitative detection of marker compounds (quercetin or gallic acid).

• Quantitative estimation of bioactive constituents by Spectrophotometric methods;

1. 1. 1. Estimation of total polyphenolics (gallic acid equivalent) in phytoextract
      2. Estimation of total flavonoids (quercetin equivalent) in phytoextract

• HPLC Analysis of phytochemical extracts;

1. 1. 1. HPLC Machine overview
      2. Sample preparation for HPLC analysis
      3. Preparation of standard plot of standard compound/marker
      4. Detection and quantification of marker compound in test samples

• Biological activity of Plant Extracts;

1. 1. In vitro anti-microbial activity 
   2. In vitro anti-inflammatory activity
   3. In vitro anti-oxidant activity

Plant Biotechnology: Molecular Detection of Plant Diseases – Advance

Basic Module 14

Duration: 20-30 Days

Training fees: ` 15000/-

Course Content

• Basics of Lab Safety: Lab safety and Procedures, Scope & Applications, Record Maintenance, Handling of Equipment

• Overview and types of Sterilization techniques, Concept of Solution Chemistry and Concentration, Keeping & storage of lab chemicals, reagents & solvents.

• Introduction to Plant Diseases and Molecular Plant Pathology

1. Overview of plant pathogens (fungi, bacteria, viruses, nematodes)
2. Why molecular detection is needed (speed, accuracy, sensitivity)
3. Comparison: conventional vs. molecular diagnostics

• Molecular lab workflow, Contamination control, use of PPE, Nuclease-free techniques and sterile handling.

• DNA / RNA Extraction & Sample Preparation

1. 1. 1. Sample Collection & Storage:
         1. Collection of infected leaves, stems, fruits, roots
         2. Transport, labeling & preservation
         3. Use of cold chain & sample documentation
      2. Genomic DNA / RNA Extraction
         1. Extraction from plant tissue, fungal mycelium, bacterial colonies
         2. CTAB method, commercial kits
         3. Pellet washing, purity check and storage

• PCR Basics, Primer Design & PCR Amplification

• Components of PCR reaction

• Pathogen-specific primers (ITS/fungal, 16S/bacteria, coat protein/virus)

• Positive, negative and no-template controls

• PCR Amplification (for any one type of disease)

• Preparing master mix in sterile environment

• Thermal cycler operation (denaturation–annealing–extension)

• Troubleshooting failed amplifications

• Agarose Gel Electrophoresis

• Gel preparation (buffer, agarose %, ethidium bromide / safer dyes)

• Loading samples & markers

• Band visualization under UV gel documentation system

• Data Analysis & Reporting

• Preparing molecular diagnostic report

• Linking molecular results with disease symptoms

• Photograph and documentation standards

Plant Biotechnology: Genetic Diversity Study of Plants– Advance

Basic Module 15

Duration: 20-30 Days

Training fees: ` 15000/-

Course Content

• Basics of Lab Safety: Lab safety and Procedures, Scope & Applications in Genetic Research, Record Maintenance, Notebook Formats, Handling of Equipment

• Overview and types of Sterilization techniques, Concept of Solution Chemistry, pH and Concentration, Keeping & storage of lab chemicals, reagents & solvents, DNA/RNA contamination control.

• Introduction to Plant Genetic Diversity

• Importance of genetic variation in crop improvement
• Molecular markers vs. morphological markers
• Types of markers: RAPD, ISSR, SSR, AFLP, SNP
• Applications: species identification, phylogeny, population diversity analysis

• Molecular Lab Workflow

• Nuclease-free techniques
• Use of PPE, pipetting accuracy
• Sterile handling practices for DNA-based experiments
• Avoiding PCR contamination (template & aerosol issues)

• DNA Extraction & Sample Preparation

• Collection of fresh and dried leaves
• Transport, labelling, barcoding & voucher specimen filing
• Preservation: silica gel, cold storage, ethanol storage
• Genomic DNA extraction by CTAB method from plant tissue
• Handling polysaccharide- and phenolic-rich samples
• Pellet washing, RNase treatment
• DNA purity check and long-term storage

• PCR for Genetic Diversity

• PCR Basics & Primer Selection: RAPD, ISSR, SSR primers (any one)
• Positive, negative and no-template controls
• PCR Master mix preparation
• Optimization of annealing temperature
• Running PCR for diversity marker sets
• Troubleshooting (smear, no band, faint band)

• Gel Electrophoresis & Band Scoring

• Preparing agarose gel (1%–2%), loading buffer, DNA ladder
• Visualization under UV gel documentation system
• Clear band scoring for RAPD, ISSR, SSR fingerprints
• Data tabulation of polymorphic / monomorphic bands

• Genetic Data Analysis

• Binary scoring (1/0) matrix preparation
• Polymorphic Information Content (PIC)
• Cluster analysis using dendrograms
• Interpretation: similarity, genetic distance and diversity

• Software Tools (Introduction & Demonstration)

• NTSYSpc / DARwin / MEGA software
• UPGMA tree generation
• Band similarity index and population structure basics

• Final Report Preparation

• Preparation of genetic diversity report
• Photograph and documentation standards
• Table & dendrogram representation
• Linking genetic results with plant traits or locality variation

Animal Biotechnology: Animal Tissue Culture Technique – Advance

Module 16

Duration: 20-30 Days

Number of seats per batch: 10 only

Training fees: ` 25000/-

Course Content

• Concept of Animal Cell & Tissue Culture, Scope & Applications.

• Basics of Lab Safety: Overview of cell culture laboratory, Lab safety and Procedures, Record maintenance, Equipments used in cell culture lab

• Aseptic handling techniques for handling cell lines and related procedures

• Detection of contaminants in cell culture and control measures

• Types of animal cells and their characteristics in culture

• Preparation of cell culture media and their sterilization methods

• Revival of cells from Liquid nitrogen (cell bank)

• Sub-culturing or passaging of cells

• Inverted microscopy of cultured cells, Cell counting and cell viability test

• Cryopreservation of cell lines

• Cytotoxic activity of chemical or drug on cell lines

1. 1. Preparation of test drug stock and control for study
   2. Dilution of test samples
   3. Revival and seeding of mammalian cells
   4. Treatment of cells with different concentrations of test drug or chemical samples
   5. Assessment of cell cytotoxicity or cellular activity by MTT assay
   6. Determination of IC50 values of test samples against the cell line

Animal Biotechnology: CAM Assay in Chick Embryo Model – Advance

Basic Module 17

Duration: 30-45 Days

Number of seats per batch: 10 only

Training fees: ` 20000/-

Course Content

• Basics of Lab Safety: Overview of Animal biosafety guidelines and ethical considerations, PPE usage, sterile handling, disinfection, Lab safety and Procedures, Record maintenance & experimental documentation, Equipments, Incubator and egg-handling safety used in lab.

• Introduction to CAM (Chorioallantoic Membrane) Assay

1. • Concept of Chick Embryo Model in Pharmacological Studies, Scope & Applications

• Anatomy of chick embryo and CAM development stage
• Advantages of CAM model for angiogenesis, anti-cancer, and anti-angiogenic studies
• Applications in drug discovery, nanomaterials testing, plant extracts & biomedical research

• Egg Selection, Storage & Incubation

• Fertile egg quality check and acquisition
• Storage, sterilization of eggshell
• Incubation conditions: temperature, humidity, turning schedule
• Day-wise embryo developmental stages

• Windowing of Eggs

• Aseptic preparation of egg surface
• Creating a window for CAM exposure
• Sealing methods using parafilm / tape
• Troubleshooting: embryo mortality, contamination, bleeding

• Preparation of Test Samples

1. 1. • Plant extract / nanoparticle / drug formulation preparation
      • Concentration and sterile filtration
      • Positive and negative controls
      • Dose preparation and application strategy

• CAM Assay Experimental Setup

1. 1. • Direct application on CAM
      • Disc method / gel sponge method
      • Incubation and observation timeline

• Monitoring Angiogenesis

1. 1. • Observation of blood vessel formation
      • Image capturing techniques
      • Endpoints: branching, vessel number, vessel length, tortuosity
      • Scoring methods for angiogenesis / anti-angiogenesis

• Data Collection & Image Analysis

1. 1. • Digital microscopy & photographic documentation
      • ImageJ/AngioTool software for vessel quantification
      • Graph plotting and statistical analysis basics

• Result Interpretation & Report Writing

1. • Comparative evaluation of treated vs control
   • Error sources and troubleshooting
   • Report structure: objective → method → results → conclusion
   • Graphs, tables, micrographs & scoring sheet

Animal Biotechnology: Pharmacological Studies Using Drosophila Model – Advance 

Basic Module 18

Duration: 60 Days

Number of seats per batch: 10 only

Training fees: ` 20000/-

Course Content

• Basics of Lab Safety: Overview of Animal biosafety guidelines and ethical considerations, PPE usage, sterile handling, disinfection, Lab safety and Procedures, Record maintenance & experimental documentation, Equipment, Handling glassware, CO₂ anaesthesia used in Fly Lab.

• Introduction to Drosophila as a Model Organism

1. 1. • Why Drosophila is used in genetics, toxicology, developmental biology
      • Life cycle: egg → larva → pupa → adult
      • Male vs female identification
      • Strains and wild-type vs mutant types

• Introduction to Pharmacological Studies in Drosophila

• Overview of D. melanogaster as a pharmacological model

• Genetic advantages, lifespan, cost-effectiveness

• Ethics & biosafety compliance in working with Drosophila

• Laboratory Setup for Drug Testing

• Stock maintenance of wild-type & mutant strains

• Preparation of fly food media with additives

• Dose preparation: solvents, stock solutions, serial dilution

•  Drug Administration Methods

• Oral administration via food supplementation

• Soaking method

• Microinjection basics

• Advantages & limitations of each method

• Pre- Experimental Handling

• Synchronization of flies (age-matched)

• Sex sorting, anaesthesia techniques (CO₂, ice)

• Designing control vs treated groups

• Replication and proper sample size

• Toxicity Assays

1. 1. • Acute toxicity (24–48 hr)
      • Chronic toxicity (lifespan alteration)
      • Survival curves (Kaplan–Meier plot demonstration)

• Behavioural Pharmacology

• Locomotor activity assay

• Climbing assay (Negative geotaxis test)

• Sleep/activity monitoring basics

• Oxidative Stress Assays

1. 1. • Paraquat/H₂O₂ induced stress
      • Survivability studies
      • ROS estimation (DHE staining demonstration)

•  Biochemical Analysis

• Protein extraction from flies

• Enzyme-based assays: Catalase, SOD, GST

• Data recording & normalization

• Reproductive Toxicity Assay

• Fecundity & fertility counting

• Hatchability % determination

• Gonad dissection basics

• Genotoxicity Assay

• SMART test (Somatic Mutation and Recombination Test)

• Chromosomal aberration basics

• Histopathological Examination

• Tissue extraction (midgut, brain, malpighian tubules)

• Fixation, staining and slide preparation

• Microscopic observations & scoring

• Dose–Response & IC50 Determination

• Calculating LC50/IC50 values

• Graph plotting & curve fitting

• Use of software (Origin/GraphPad)

• Data Interpretation & Statistics

• Mean, SD, SEM

• Student t-test, ANOVA

• Representation: tables, graphs, significance values

• Experimental Report Preparation

• Standard format: Introduction–Method–Results–Conclusion

• Photograph & graph presentation

• Common mistakes & troubleshooting

• Data recording and documentation