Laboratory Protocols in Applied Life Sciences

<P><STRONG>Microscopy</STRONG></P>
<P>Introduction</P>
<P>Light Microscopy</P>
<P>Atomic Force Microscopy</P>
<P>Bright Field Microscopy</P>
<P>Dark Ground Microscopy</P>
<P>Phase Contrast Microscopy</P>
<P>Confocal Scanning Laser Microscopy</P>
<P>Differential Interference Contrast Microscopy</P>
<P>Polarization Microscopy</P>
<P>Fluorescence Microscopy</P>
<P>Experiments on Microscopy</P>
<P>Electron Microscopy</P>
<P>Sample Preparation for Electron Microscopy</P>
<P>Dark Field Electron Microscopy</P>
<P>Crewe Microscope</P>
<P>Backscatter Scanning Microscope</P>
<P>Experiment on Scanning Electron Microscopy of Cyanobacteria</P>
<P>Transmission Electron Microscopy of Phages (Bacteriophage and Cyanophage)</P>
<P>Suggested Readings</P>
<P>Important Links</P><STRONG>
<P>Microtome</P></STRONG>
<P>Introduction</P>
<P>Sectioning</P>
<P>Traditional Histology</P>
<P>Ultramicrotome</P>
<P>Cryostat</P>
<P>Incubation</P>
<P>Electron Microscopy Technique</P>
<P>Botanical Microtomy</P>
<P>Processing of Tissue</P>
<P>Suggested Readings</P>
<P>Important Links</P><STRONG>
<P>pH and Buffers</P></STRONG>
<P>Introduction</P>
<P>Acids and Bases</P>
<P>Hydrogen Ion Concentration and pH</P>
<P>Measurement of pH</P>
<P>Titration Curves</P>
<P>Buffer Solutions</P>
<P>What Is the pH of a Mixture of 5 mL of 0.1 mol/L Sodium Acetate and 4 mL of 0.1 mol/L Acetic Acid?</P>
<P>How the pH Changed on Adding 1 mL of 0.1 mol/L HCl in the Previous Mixture?</P>
<P>Buffers Used in Biochemical Experiments</P>
<P>pH and Life</P>
<P>Care and Use of the pH Meter</P>
<P>Titration Curves</P>
<P>Determination of pH Using Indicators</P>
<P>Titration of a Mixture of a Strong and a Weak Acid</P>
<P>Titration of a Strong Acid with a Strong Base</P>
<P>Titration of a Weak Acid with a Strong Base</P>
<P>Determination of pKa</P>
<P>pKa Values of a Dicarboxylic Acid</P>
<P>Acetate Buffers</P>
<P>Titration Curves of Amino Acids</P>
<P>Determination of pH of the Given Water/Soil Sample</P>
<P>Determination of pH of Fruit Juice</P>
<P>Measurement of pH of Soil Sample</P>
<P>Appendices</P>
<P>Suggested Readings</P>
<P>Important Links</P><STRONG>
<P>Spectrophotometer</P></STRONG>
<P>Introduction</P>
<P>Beer-Lambert Law</P>
<P>Limitations of the Beer-Lambert Law</P>
<P>Measurement of Extinction</P>
<P>Colorimeter</P>
<P>Spectrophotometry</P>
<P>Infrared Spectroscopy</P>
<P>Fluorescence Spectroscopy</P>
<P>Sensitivity of Fluorescence Assays</P>
<P>Fluorescence Quenching</P>
<P>Measurement of ?-Naphthyl Phosphatase Activity</P>
<P>Suggested Readings</P>
<P>Important Links</P><STRONG>
<P>Centrifugation</P></STRONG>
<P>Introduction</P>
<P>Centrifuges</P>
<P>Ultracentrifugation</P>
<P>Suspending Medium</P>
<P>Rotors</P>
<P>Fractionation of Rat Liver</P>
<P>Estimation of DNA, RNA, and Protein in the Isolated Cell Fractions</P>
<P>Enzyme Distribution in the Cell</P>
<P>Glucose-6-Phosphatase</P>
<P>Glutamate Dehydrogenase</P>
<P>Acid Phosphatase</P>
<P>Mitochondrial Swelling</P>
<P>Determination of Lysosomal Integrity</P>
<P>Effect of Detergents on the Stability of the Lysosomal Membrane</P>
<P>Density Gradient Centrifugation</P>
<P>Fractionation of Pig Brain by Density Gradient Centrifugation</P>
<P>Distribution of Enzymes in the Fractions Obtained from Pig Brain</P>
<P>Isolation of Chloroplasts</P>
<P>Isolation of Chloroplast DNA</P>
<P>Isolation of Mitochondria and Assay of a Marker Enzyme</P>
<P>Isolation of Genomic DNA</P>
<P>Isolation of Chloroplasts from Spinach Leaves by Differential Centrifugation</P>
<P>Isolation of Plasmid DNA by Centrifugation</P>
<P>Suggested Readings</P>
<P>Important Links</P><STRONG>
<P>Radiation Biology</P></STRONG>
<P>Introduction</P>
<P>Radioisotopes</P>
<P>Sample Preparation</P>
<P>Autoradiography</P>
<P>Distribution of <SUP>14</SUP>CO<SUB>2</SUB> in Different Plant Parts </P>
<P>Photosynthetic Reduction of <SUP>14</SUP>CO<SUB>2</SUB> to Primary Metabolic Product</P>
<P>Incorporation of Labeled Acetate into Leucoplastic Fatty Acids of Developing Brassica Seeds</P>
<P>Suggested Readings</P>
<P>Important Links</P><STRONG>
<P>Chromatography</P></STRONG>
<P>Chromatography</P>
<P>Paper Chromatography</P>
<P>Ascending Chromatography</P>
<P>Descending Chromatography</P>
<P>Separation and Identification of Amino Acids by Descending Paper Chromatography</P>
<P>Separation and Identification of Amino Acids by Ascending Paper Chromatography</P>
<P>Separation and Identification of Amino Acids in a Given Mixture by 2 D Paper Chromatography</P>
<P>Thin Layer Chromatography</P>
<P>Thin-Layer Gel Chromatography</P>
<P>Separation and Identification of Sugars by Adsorption Thin Layer Chromatography</P>
<P>Identification of Lipids in a Given Sample by Thin Layer Chromatography</P>
<P>Identification of Sugars in Fruit Juices Using Thin Layer Chromatography</P>
<P>Column Chromatography</P>
<P>Gradient Preparation</P>
<P>Adsorption Chromatography</P>
<P>Separation of Pigments from Leaves or Flowers by Adsorption Column Chromatography</P>
<P>Ion-Exchange Chromatography</P>
<P>Separation of Amino Acids by Ion-Exchange Chromatography</P>
<P>Separation of Proteins from Human Serum by Ion-Exchange Chromatography</P>
<P>Gel Filtration/Size-Exclusion Column Chromatography</P>
<P>Separation of Blue Dextran and Cobalt Chloride on Sephadex G-25</P>
<P>Desalting of Protein Sample by Gel Filtration</P>
<P>Determination of Molecular Weight of a Given Protein by Gel Filtration</P>
<P>Concentration of Dilute Protein Solutions Using Sephadex G-25</P>
<P>Affinity Chromatography</P>
<P>Hydrophobic Interaction Chromatography or Affinity Elution</P>
<P>Purification by Affinity Chromatography</P>
<P>Preparation of an Affinity Column</P>
<P>Preparation of Rat Brain Acetylcholinesterase</P>
<P>Acetylcholinesterase Assay</P>
<P>Purification of Acetylcholinesterase by Affinity Chromatography</P>
<P>MALDI-TOF</P>
<P>Nuclear Magnetic Resonance Spectroscopy</P>
<P>Maintenance of Enzyme Activity</P>
<P>Addition of Chaotropes</P>
<P>Role of Polyols/Hydrophilic Polymers in Stabilizing Structures</P>
<P>Stabilization by Side-Chain Modification</P>
<P>Criteria of Purity</P>
<P>Biuret</P>
<P>Folin (Lowry's Method)</P>
<P>Bradford</P>
<P>Suggested Readings</P>
<P>Important Links</P><STRONG>
<P>Separation Technology</P></STRONG>
<P>Electrophoresis</P>
<P>Principle</P>
<P>Types of Electrophoresis</P>
<P>Separation of Amino Acids by Paper Electrophoresis</P>
<P>Cellulose Acetate Strip Electrophoresis</P>
<P>Gel Electrophoresis</P>
<P>Polyacrylamide Gel Electrophoresis</P>
<P>Polyacrylamide Disc Gel Electrophoresis</P>
<P>SDS Gel Electrophoresis</P>
<P>Slab Gel Electrophoresis</P>
<P>Native Disc Gel Electrophoresis of Proteins</P>
<P>SDS-Polyacrylamide Slab Gel Electrophoresis of Proteins Under Reducing Conditions (SDS-PAGE)</P>
<P>Slab Gel Gradient Electrophoresis of Protein and Multiple Molecular Forms of Acetylcholinesterase</P>
<P>Protein (Western) Blotting</P>
<P>Isoelectric Focusing</P>
<P>Agarose Gel Electrophoresis</P>
<P>Determination of Molecular Weight of Plasmid DNA by Agarose Gel Electrophoresis</P>
<P>Electrophoretic Evaluation of Quality of High Molecular Weight DNA Isolated from Plant Material</P>
<P>Large-Scale Preparative Electrophoresis</P>
<P>Suggested Readings</P>
<P>Important Links</P><STRONG>
<P>Histochemical Techniques</P></STRONG>
<P>Introduction</P>
<P>Protein</P>
<P>Acidic Compounds</P>
<P>Carbohydrates-Polysaccharides</P>
<P>Lipids</P>
<P>Minor Compounds</P>
<P>Autoradiography</P>
<P>Enzyme Localization in Embedded Material</P>
<P>Suggested Readings</P>
<P>Important Links</P><STRONG>
<P>Cytogenetics</P></STRONG>
<P>Introduction</P>
<P>Study Different Stages in Root Tip Cells of Plant</P>
<P>Preparation of Root Tips for Mitotic Studies</P>
<P>Preparation and Study of Slides for Mitosis Using Acetocarmine Squash Techniques from Onion Root Tips</P>
<P>Perform Carmine or Orcein Staining Techniques</P>
<P>Feulgen Staining Techniques</P>
<P>Preparation of Onion Flower Buds for Meiosis Studies</P>
<P>Fixation</P>
<P>Mounting and Making Temporary Slides Permanent</P>
<P>Study the Effect of Colchicines or Acenaphthene or Chloral Hydrate on Chromosomes of <I>Allium cepa</P></I>
<P>Estimate the Chiasma Frequency at Diplotene-Diakinesis</P>
<P>Study the Effect of ? Radiation on Chromosomes</P>
<P>To Prepare Slides Showing Diplotene Stages in Spermatocytes of Desert Locust (<I>Schistocerca gregaria</I>) or in Oocytes of the Newt (<I>Triturus viridescens</I>) Showing Lampbrush Chromosomes</P>
<P>Study Karyotypes of Allium cepa and <I>Vicia faba</P></I>
<P>Micrometry</P>
<P>To Demonstrate Reciprocal Translocation in Pollen Mother Cells of Rhoeo discolor (Tradescantia)</P>
<P>To Study the Spiral Nature of Chromosome Using Any Plant Material</P>
<P>To Show the Different Banding Patterns in Eukaryotic Chromosomes</P>
<P>Fungal Cytogenetics</P>
<P>Harvesting of Neurospora</P>
<P>To Study the Ordered (Tetrad) and Random Ascospore Analysis in Neurospora sp</P>
<P>To Demonstrate the Tetrad and Random Analysis</P>
<P>Tetrad Analysis</P>
<P>Pigmentation and Nutritional Requirement Analysis</P>
<P>Random Spore Analysis</P>
<P>Test for Nutritional Requirement</P>
<P>Suggested Media for Neurospora, Sordaria, and Aspergillus Experiments</P>
<P>Study of Heterokaryosis in Neurospora from the Nutritionally Deficient Mutants</P>
<P>General Information and Methods for Drosophila Experiments</P>
<P>Study of Salivary Gland Chromosomes of Drosophila melanogaster for Nature of Polyteny and Regions of Chromosome Puffs</P>
<P>Human Genetics</P>
<P>Human Sex Chromosomes and Barr Bodies</P>
<P>Autoradiography</P>
<P>Photomicrography</P>
<P>Suggested Readings</P>
<P>Important Links</P><STRONG>
<P>Biomolecules</P></STRONG>
<P>Introduction</P>
<P>Carbohydrates</P>
<P>Amino Acids and Proteins</P>
<P>Lipids</P>
<P>Nucleic Acids</P>
<P>Suggested Readings</P>
<P>Important Links</P><STRONG>
<P>Enzymology</P></STRONG>
<P>Introduction</P>
<P>Enzymes as Catalysts</P>
<P>Classification</P>
<P>Measurement of Enzyme Activity</P>
<P>Time Course Studies of the Reaction Catalyzed by Alkaline Phosphatase (EC 3.1.3.1)</P>
<P>Effect of Enzyme Concentration on the Rate of Enzyme-Catalyzed Reaction</P>
<P>Determination of Temperature Optima for Alkaline Phosphatase</P>
<P>Determination of Effect of pH on the Activity of Alkaline Phosphatase</P>
<P>Effect of Substrate Concentration on Activity of Alkaline Phosphate and Determination of the Km and Vmax of the Reaction</P>
<P>Progress Curve of Serum Alkaline Phosphatase (Orthophosphoric Acid Monoester Phosphohydrolase, EC 3.1.3.1)</P>
<P>Effect of Variation of Serum Alkaline Phosphatase Activity with Enzyme Concentration</P>
<P>Effect of Substrate Concentration and Inhibitors on Oxheart Lactate Dehydrogenase</P>
<P>Effect of Temperature on the Activity of ?-Amylase</P>
<P>Determination of the Molecular Weight and Purity of Chymotrypsin from the Enzyme Kinetics</P>
<P>Yeast Isocitrate Dehydrogenase: Allosteric Enzyme</P>
<P>Purification and Characterization of Enzymes</P>
<P>Isolation of Muramidase (Mucopeptide N-Acetylmuramyl Hydrolase, ) from Egg White</P>
<P>Separation of the Isoenzymes of Lactate Dehydrogenase by Electrophoresis on Polyacrylamide Gel</P>
<P>Enzyme Engineering</P>
<P>Artificial Enzymes</P>
<P>Suggested Readings</P>
<P>Important Links</P><STRONG>
<P>Membranes</P></STRONG>
<P>Introduction</P>
<P>Composition of Membranes</P>
<P>Membrane Transport</P>
<P>Effect of Lipid Composition on the Permeability of a Lipid Monolayer</P>
<P>Effect of Detergents and Other Membrane-Active Reagents on the Erythrocyte Membrane</P>
<P>Permeability of Model Membrane (Liposome)</P>
<P>Effect of Cholesterol on the Anion Permeability of a Phospholipid Membrane</P>
<P>Effect of Insulin on the Transport into Isolated Fat Cell</P>
<P>Transport of Amino Acids across the Small Intestine</P>
<P>Absorption of Xylose from the Gut in Man</P>
<P>Suggested Readings</P>
<P>Important Links</P><STRONG>
<P>Photosynthesis and Respiration</P></STRONG>
<P>Introduction</P>
<P>Measurement</P>
<P>Photosynthesis</P>
<P>Isolation of Chloroplasts from Spinach Leaves</P>
<P>Determination of Chlorophyll Concentration in the Chloroplast Suspension</P>
<P>Evolution of Oxygen by Isolated Chloroplast Using Hill Oxidants</P>
<P>Methyl Viologen as a Terminal Electron Acceptor</P>
<P>Spectrophotometric Assay of the Hill Reaction and the Estimation of Chlorophyll</P>
<P>Respiration of Mitochondria and Oxidative Phosphorylation</P>
<P>Effect of Inhibitors on the Respiratory Chain</P>
<P>Compounds That Affect the High-Energy State of Mitochondria</P>
<P>Suggested Reading</P>
<P>Important Links</P><STRONG>
<P>Microbiology</P></STRONG>
<P>Introduction</P>
<P>Good Microbiological Practice</P>
<P>Culture Media</P>
<P>Types of Media</P>
<P>Preparation of Culture Media</P>
<P>Maintenance and Preservation of Cultures</P>
<P>Preparation of Basic Liquid Media (Broth) for the Cultivation of Bacteria</P>
<P>Solid Media for Growth of Microorganisms</P>
<P>Isolation and Maintenance of Organisms</P>
<P>Isolation of Organisms by Pour Plate Method and Serial Dilation Method</P>
<P>Spread Plate Technique</P>
<P>Isolation of <I>Vibrio parahaemolyticus</I> from Seafood Sample</P>
<P>Isolation of Lactic Acid Bacteria from Milk and Buttermilk Sample</P>
<P>Isolation of <I>Staphylococcus aureus</I> from Food Sample</P>
<P>Sauerkraut Fermentation</P>
<P>Isolation of Microorganisms from Air</P>
<P>Phyllosphere/Phylloplane Microbiology</P>
<P>Measurement of Microbial Growth</P>
<P>Methods to Measure Bacterial Growth</P>
<P>Growth Curve Measurement of Bacterial Population by Serial Dilution Method</P>
<P>Serial Dilutions</P>
<P>Measurement of Fungal Growth by Colony Diameter Method</P>
<P>Biomass Measurement of Fungal Growth (Dry Weight Method)</P>
<P>Effect of Incubation Temperature on Growth</P>
<P>Effect of pH on Microbial Growth</P>
<P>Microscopic Examination of Bacteria</P>
<P>Microscopic Examination of Pathogens by One Tube One Plate Method</P>
<P>Smear Preparation</P>
<P>Staining</P>
<P>Biochemical Tests</P>
<P>Microbial Genetics</P>
<P>Bacterial Recombination</P>
<P>Suggested Readings</P>
<P>Important Links</P><STRONG>
<P>Soil Biology</P></STRONG>
<P>Introduction</P>
<P>Estimation of Humus in Soil/Manure</P>
<P>Determination of Microbial Carbon Biomass</P>
<P>Estimation of Algal (Photosynthetic) Biomass in Soil Cores</P>
<P>Soil Decomposition and Microbial Community Structure (Winogradsky Technique)</P>
<P>Soil Algae Enumeration</P>
<P>Phosphate Solubilizing Bacteria</P>
<P>Isolation of Sulfur Oxidizing Bacteria from Soil</P>
<P>Demonstrate the Process of Ammonification in Soil</P>
<P>Demonstrate the Process of Nitrification in Soil</P>
<P>MPN Method for Nitrosomonas and Nitrobacter</P>
<P>Isolation of Azospirillum sp from Soil and Plant Root</P>
<P>Enumeration of Azospirillum by MPN Method</P>
<P>Demonstration of Microbial Denitrification in Soil</P>
<P>Isolation of Plant Growth Promoting Rhizobacteria</P>
<P>Isolation of Photosynthetic Bacteria</P>
<P>Cyanobacteria</P>
<P>Nitrogen Fixation</P>
<P>Mycorrhizal Symbiotic Association</P>
<P>Actinomycetes</P>
<P>Aquatic Molds</P>
<P>Fungi</P>
<P>Rhizosphere</P>
<P>Microbial Production of Citric Acid by <I>Aspergillus niger</P></I>
<P>Isolation of Industrially Important Antibiotic Producing Microorganisms</P>
<P>Assay of Antibiotics and Demonstration of Antibiotic Resistance</P>
<P>Characterize and Identify Aflatoxin by Thin-Layer Chromatography</P>
<P>Protozoa</P>
<P>Observation of Microscopic Invertebrates from Pond Water</P>
<P>Observation of Algae from Pond Water</P>
<P>Microscopic Examination of Soil Microbes</P>
<P>Bacteria Frequently Isolated from Soil</P>
<P>Fungi Frequently Isolated from Soil</P>
<P>Appendices</P>
<P>Suggested Reading</P>
<P>Important Links</P><STRONG>
<P>Virology</P></STRONG>
<P>Introduction</P>
<P>Isolation of Bacteriophage from Sewage</P>
<P>Preparation of High Titer of Bacteriophage of Isolated Bacterial Virus</P>
<P>One-Step Growth Curve of Coliphage and Determining the Number of Phage Particles Released by the Lysis of the Single Infected Bacterial Cell (Burst Size)</P>
<P>Demonstration of Lysogeny and Isolation of Rhizobium Phage</P>
<P>Isolation of Cyanophage LPP from Pond Water</P>
<P>Demonstration for the Presence of Actinophages in Soil</P>
<P>Isolation of Phage from Flies</P>
<P>Virus Culture in Embryonating Chicken Eggs</P>
<P>Extraction and Isolation of Tobacco Mosaic Virus</P>
<P>Cultivation of Viruses in Tissue Culture</P>
<P>Viral Hemagglutination Test for the Presence of Antigens</P>
<P>Suggested Readings</P><STRONG>
<P>Immunochemical Methods</P></STRONG>
<P>Introduction</P>
<P>Immunity</P>
<P>Ags</P>
<P>Antigenic Determinant or Epitope</P>
<P>Haptens</P>
<P>Specificity</P>
<P>T-Cell Dependent (TD) and T-Cell Independent (TI) Ags</P>
<P>Immunoglobulins</P>
<P>IG Determinants</P>
<P>Ig Classes</P>
<P>Monoclonal Antibodies</P>
<P>Polyclonal Antibodies</P>
<P>Ag-Ab Reaction</P>
<P>Determination of Ag-Ab Reactions</P>
<P>Collection of Blood Sample</P>
<P>Experiment: To Determine the ABO Blood Group by Using Slide Agglutination Test</P>
<P>Experiment: Determination of RH Factor in Human Being</P>
<P>Experiment: Demonstration of Agglutination Reaction of Unknown Bacterial Culture by Slide Agglutination Technique</P>
<P>Experiment: Estimate the Total Leukocyte Count by Hemocytometer</P>
<P>Experiment: Determine the Differential Count of WBCs (Leucocyte) by Staining Method</P>
<P>Experiment: Determine the Total Erythrocyte Count by Hemocytometer</P>
<P>Experiment: Lymphoid Cells Identification in Blood Smears</P>
<P>Experiment: Spleen Cell Preparation from Mouse</P>
<P>Experiment: Identification of Lymphocyte Populations</P>
<P>Experiment: Isolation of Monocytes by Adherence</P>
<P>Experiment: Isolation of Neutrophils</P>
<P>Experiment: Isolation of Human Basophils</P>
<P>Experiment: Isolation of Tissue Mast Cells</P>
<P>Experiment: Measurement of Percent ?-Glucuronidase Secretion</P>
<P>Experiment: Generation of Polyclonal Antibodies in Rabbits</P>
<P>Experiment: Gel Diffusion/Ouchterlony Immunodiffusion</P>
<P>Experiment: Demonstration of Radial Immunodiffusion</P>
<P>Experiment: Counter Current Immunoelectrophoresis</P>
<P>Experiment: Serum Electrophoresis</P>
<P>Experiment: Demonstration of Immunoelectrophoresis</P>
<P>Experiment: Determination of Albumin by Laurell Rocket Immunoelectrophoresis</P>
<P>Experiment: Purification of Igs BY ION Exchange Chromatography</P>
<P>Experiment: The Detection and Assay of a Myeloma Protein</P>
<P>Experiment: ELISA</P>
<P>Experiment: Pregnancy Testing by Using the Immunological Methods</P>
<P>Experiment: HIV DOT Test of AIDS Patients</P>
<P>Experiment: Demonstration of Immunofluorescence Assay</P>
<P>Experiment: Identification of Bacteria by Using Fluorescent Ab Technique (FAT)</P>
<P>Experiment: Immunofluorescence Labeling of Cultured Cells</P>
<P>Experiment: Isolation of IgG from Serum</P>
<P>Experiment: Purification of IgM</P>
<P>Suggested Reading</P>
<P>Important Links</P><STRONG>
<P>Genetic Engineering</P></STRONG>
<P>Introduction</P>
<P>Tools of Genetic Engineering</P>
<P>DNA Sequencing</P>
<P>Isolation of Genomic DNA from <I>E coli</P></I>
<P>Rapid Method for Isolating Plasmid DNA from Yeast</P>
<P>Isolation of Plasmid DNA from Yeast (Protocol 1)</P>
<P>Isolation of Yeast Plasmid DNA (Protocol 2)</P>
<P>Isolation of Drosophila Genomic DNA</P>
<P>Determination of ?max Value of the Isolated DNA Sample</P>
<P>Determination of the Purity of the Isolated DNA Sample</P>
<P>Determine the Quantity of DNA in the Sample</P>
<P>To Perform Agarose Gel Electrophoresis for the Given DNA Sample</P>
<P>Isolation of Plasmid DNA from Given Bacterial Culture</P>
<P>Electrophoresis of DNA: Linear, Circular, and Super Coiled</P>
<P>To Perform Restriction Digestion of the Given DNA Sample</P>
<P>To Demonstrate Ligation</P>
<P>Elution of DNA from Agarose Gel</P>
<P>Purification of Phage ? DNA</P>
<P>To Perform Restriction Digestion of Plasmid DNA with Different Restriction Endonucleases and to Determine the Position of Restriction Site Using Restriction Mapping</P>
<P>To Perform Transformation</P>
<P>Directional Cloning Into Plasmid Vectors</P>
<P>To Assay Reporter Chloramphenicol Acetyltransferase Gene</P>
<P>To Assay ss-Gal in Extracts of Mammalian Cells</P>
<P>Determination of Nucleotide Sequence of DNA by Dideoxy Chain Termination Method</P>
<P>Amplification of 16S rDNA Genes by the Polymerase Chain</P>
<P>PCR from Fungal Spores/AMF (Arbuscular Mycorrhizal Fungi)</P>
<P>To Demonstrate the Nested PCR Reaction</P>
<P>Suggested Readings</P>
<P>Important Links</P><STRONG>
<P>Molecular Biology</P></STRONG>
<P>Introduction</P>
<P>Isolation of Nucleic Acids from Biological Samples</P>
<P>Demonstration of Southern Blotting</P>
<P>Demonstration of Experiment on RFLP Analysis</P>
<P>Restriction Fragment Length Polymorphism</P>
<P>Isolation of RNA from Biological Sample</P>
<P>Extraction of Polysomal RNA of Nuclear Sequences</P>
<P>Separation of RNA Species by Ion-Exchange Column Chromatography on Methylated Albumin Kiesselguher Columns</P>
<P>Northern Hybridization of RNA Fractionated by Agarose Formaldehyde Gel Electrophoresis</P>
<P>Preparation of Radiolabeled Probe by Random Primer Method</P>
<P>Detection of Specific Fragment of Plant DNA from Restriction Digest by Hybridization with a Labeled Probe</P><I>
<P>In Vitro</I> Transcription</P><I>
<P>In Vitro</I> Translation of RNA</P>
<P>Reticulocytes and Their Use in the Study of Protein Synthesis</P>
<P>Protein Synthesis in Intact Rabbit Reticulocytes</P>
<P>Metabolic Labeling of Proteins and Immunoprecipitation</P>
<P>Induction of ss-Galactosidase in Strains of <I>E coli</P></I>
<P>Effect of Different Inducers on the Induction of ss-Galactosidase</P>
<P>Effect of Protein Synthesis Inhibitors on the Induction of ss-Galactosidase</P>
<P>Turnover of ss-Galactosidase</P>
<P>Suggested Readings</P>
<P>Important Links</P><STRONG>
<P>Biosensors</P></STRONG>
<P>Introduction</P>
<P>Biosensor</P>
<P>Classification of Biosensors</P>
<P>Characteristics of Biosensors</P>
<P>Components of Biosensors</P>
<P>Biochip</P>
<P>Immunobiosensor</P>
<P>Aptamers</P>
<P>SPR Technology</P>
<P>Working Principle of Biosensors</P>
<P>Immunosensors</P>
<P>Calorimetric Biosensors</P>
<P>Potentiometric Biosensors</P>
<P>Amperometric Biosensors</P>
<P>Optical Biosensors</P>
<P>Piezoelectric Biosensors</P>
<P>Second-Generation Biosensors</P>
<P>Third-Generation Biosensors</P>
<P>Biosensor for Healthcare</P>
<P>Conclusion and Future Challenges</P>
<P>Suggested Readings</P>
<P>Important Links</P><STRONG>
<P>Enzyme Immobilization</P></STRONG>
<P>Introduction</P>
<P>Selection of Carrier and Support before Immobilization</P>
<P>Carriers for Enzyme Immobilization</P>
<P>Limitations and Advantages of Enzyme Immobilization</P>
<P>Methods of Immobilizations</P>
<P>Cross-Linking</P>
<P>Entrapment Method</P>
<P>Whole-Cell Immobilization</P>
<P>Industrial Application of Enzymes</P>
<P>Experiment Cell Immobilization by Using Alginate</P>
<P>Experiment Immobilization of Enzyme Horseradish</P>
<P>Peroxidase (Donor: H<SUB>2</SUB>O<SUB>2</SUB>Oxidoreductase; EC 1.11.1.7) by</P>
<P>Aminoantipyrine-Phenol Assay</P>
<P>Suggested Readings</P><STRONG>
<P>Plant Tissue Culture</P></STRONG>
<P>Introduction</P>
<P>Facilities</P>
<P>Glassware Preparation</P>
<P>Sterilization</P>
<P>Wet Heat Sterilization</P>
<P>Dry Sterilization</P>
<P>Filter Sterilization</P>
<P>Media Preparation</P>
<P>Preparation of Stock Solution</P>
<P>Protocol</P>
<P>Sterilization of Media</P>
<P>Aseptic Manipulation</P>
<P>Sterile Transfer Facilities</P>
<P>Temperature</P>
<P>Light</P>
<P>Aeration</P>
<P>Temperature</P>
<P>Plant Cell Culture System</P>
<P>Explants</P>
<P>Calluses</P>
<P>Cell Suspension</P>
<P>Protoplast</P>
<P>Surface Sterilization</P>
<P>Common Culture Media</P>
<P>Sterilization of Plant Materials</P>
<P>Introduction</P>
<P>Materials</P>
<P>Protocol</P>
<P>Observation and Result</P>
<P>Preparation of Tissue Culture Media</P>
<P>Organ Culture</P>
<P>Callus Propagation</P>
<P>Single Cell Culture</P>
<P>Preparation and Fusion of Protoplasts</P>
<P>Protocols on Protoplast Fusion</P>
<P>Fusion Products: Hybrids and Cybrids</P>
<P>Methods of Somatic Hybridization</P>
<P>Protoplast Fusion (Somatic Cell Hybridization)</P>
<P>Electrofusion Isolation of Oat and Corn Protoplast</P>
<P>Culture of Protoplasts</P>
<P>Other Protocols</P>
<P>Somatic Embryogenesis</P>
<P>Production of Haploid Plants</P>
<P>Embryo Encapsulation: Production of Artificial Seeds</P>
<P>Cytological Examination of Regenerated Plants</P>
<P>Demonstration of Root Hair Culture of Different Explants Source of Cereal Using <I>Agrobacterium rhizogenes</P></I>
<P>Experiment Demonstration of Root Hair Culture of Different Explants Using Arbuscular Mycorrhizal Fungi</P>
<P>Useful Secondary-Metabolite Production from Plant Tissue Culture</P>
<P>Genetic Transformation of Plants through <I>Agrobacterium tumefaciens</I> as Cloning Vector</P>
<P>Assay of ss-Glucuronidase in Transformed Plant Tissue</P>
<P>Random Amplified Polymorphic DNA Analysis</P>
<P>Efficiency of Rhizobia for Nodulation in Leguminous Crops</P>
<P>Seed Inoculation with Rhizobia</P>
<P>Suggested Readings</P>
<P>Important Links</P><STRONG>
<P>Animal Cell Science and Technology</P></STRONG>
<P>Introduction</P>
<P>Chronology of Cell Culture Development</P>
<P>Cell Culture Laboratory</P>
<P>Culture Medium</P>
<P>Cell Subculturing</P>
<P>Cell Quantification</P>
<P>Seeding Cells onto Culture Plates</P>
<P>Maintenance of Cells in Culture</P>
<P>Growth Kinetics of Animal Cells in Culture</P>
<P>Cryopreservation of Cells</P>
<P>Resuscitation of Frozen Cells</P>
<P>Determination of Cell Viability</P>
<P>Safety Considerations in Cell Culture</P>
<P>Aseptic Techniques and Good Cell Culture Practice</P>
<P>Preparation of Tissue Culture Medium</P>
<P>Preparation of Single Cell Suspension of Spleen and Thymus</P>
<P>Cell Counting</P>
<P>Cell Viability Assay</P>
<P>Estimation of Viability by Dye Uptake</P>
<P>Macrophage Monolayer from PEE and Measurement of Phagocytic Activity</P>
<P>Trypsinization of Monolayer and Subculturing</P>
<P>Cryopreservation</P>
<P>Thawing</P>
<P>Role of Serum in Cell Culture</P>
<P>Metaphase Chromosome Preparation from Cultured Cells</P>
<P>Isolation of DNA and Demonstration of Apoptosis of DNA Laddering</P>
<P>Demonstration of Apoptosis of DNA Laddering</P>
<P>MTT Assay for Cell Viability and Growth</P>
<P>Cell Fusion by PEG</P>
<P>Suggested Readings</P>
<P>Important Links</P><STRONG>
<P>Bioprocess Engineering</P></STRONG>
<P>Introduction</P>
<P>Isolation of Industrially Important Microorganisms</P>
<P>Determination of Thermal Death Point and Thermal Death Time of Microorganisms for Design of a Sterilizer</P>
<P>Determination of Ethanol Production Using Different Substrates</P>
<P>Sugar Fermentation by Yeast (<I>S. cerevisiae</I>) for Production of Ethanol and Cell Biomass Using Laboratory Fermenter of 2 L capacityMeasurement of Yeast Biomass Using Hemocytometer Technique</P>
<P>Study Fermentation Process for Lactic Acid Production by <I>Streptococcus thermophilus</I> and Cell Biomass</P>
<P>Microbial Production of Citric Acid Using <I>Aspergillus niger</P></I>
<P>Microbial Production of Antibiotics (Penicillin)</P>
<P>Production of Vinegar (Acetic Acid)</P>
<P>Demonstration of Wine Production Using Grape Wine Juice</P>
<P>Production and Manufacture of Beer</P>
<P>Production and Estimation of Alkaline Protease</P>
<P>Mushroom Production Technology</P>
<P>Identification of Mushrooms by Spore Print Method</P>
<P>Isolation and Maintenance of Mushroom Culture</P>
<P>Production of Spawn for White Button Mushroom (A. brunnescens syn A.bisporus)</P>
<P>Cultivation of White Button Mushroom (<I>A. brunnescens = A. bisporus</I>)</P>
<P>Cultivation of <I>G. lucidum</I> (Ling Zhi)</P>
<P>Cultivation of <I>L. edodes</P></I>
<P>Cultivation of Paddy Straw Mushroom (<I>V. volvacea</I>)</P>
<P>Cultivation Method of Oyster Mushroom</P><STRONG>
<P>Environmental Biology</P></STRONG>
<P>Wastewater Treatment Process</P>
<P>Measurement of pH</P>
<P>Measurement of Alkalinity of Water</P>
<P>Measurement of DO of Water</P>
<P>Measurement of Chloride in Water</P>
<P>Measurement of Nitrate in Water</P>
<P>Measurement of Nitrite in Water</P>
<P>Measurement of Ammonia in Water</P>
<P>Measurement of Total Phosphorus in Water</P>
<P>Measurement of Inorganic Phosphate in Water</P>
<P>Measurement of Sulfate in Water</P>
<P>Measurement of Sulfide in Water</P>
<P>Measurement of Calcium and Magnesium in Water</P>
<P>Detection of Coliforms for the Determination of the Purity of Potable Water</P>
<P>Determination of Total Dissolved Solids of Water</P>
<P>Determination of DO Concentration of Water by Winkler's Iodometric Method</P>
<P>Determination of Biochemical Oxygen Demand of Sewage Water</P>
<P>Determination Chemical Oxygen Demand Sewage Sample</P>
<P>Determination of the Efficiency of Removal of Air Pollutant, Using Fibrous Air Filter</P>
<P>Isolation of Xenobiotic-Degrading Bacteria</P>
<P>Degradation of Aromatic Hydrocarbons by Bacteria</P>
<P>Survey of Degradative Plasmids in Microbes Growing in Polluted Environment</P>
<P>Effect of SO<SUB>2</SUB> on Crop Plants</P>
<P>Estimation of Heavy Metals in Water/Soil by Atomic Absorption Spectrophotometry</P>
<P>Preliminary Digestion for Metals</P>
<P>Estimation of Metals</P>
<P>Flame AAS</P>
<P>Measurement of Aluminum in Soil/Water Sample</P>
<P>Measurement of Arsenic in Soil/Water Sample</P>
<P>Measurement of Cadmium in Soil/Water Sample</P>
<P>Measurement of Mercury in Soil/Water Sample</P>
<P>Study on Biogenic Methane Production in Different Habitats (Biogas)</P>
<P>Suggested Readings</P>
<P>Important Links</P><STRONG>
<P>Bioinformatics</P></STRONG>
<P>Introduction</P>
<P>Development</P>
<P>Nucleotide Sequence Databases</P>
<P>Protein Sequence Databases</P>
<P>Biomolecular Structure Databases</P>
<P>Pair-Wise Sequence Alignment</P>
<P>Detection of CpG Island for Nucleotide Sequence</P>
<P>Detection of Gene Structure of DNA Sequence</P>
<P>Translation of Nucleotide Sequence to Protein Sequence</P>
<P>Reports of Various Chemical and Physical Properties of Protein Sequence</P>
<P>Database Search "Blast"</P>
<P>Phylogenetic Analysis</P>
<P>Molecular Pathways Database</P>
<P>Proteomics</P>
<P>Metabolomics</P>
<P>Genomics</P>
<P>Bioinformatics Approaches for DNA-Based Signatures of Species/Populations/Breeds/Races/Variety/Strains</P>
<P>DNA-Based Signature of Plant Variety (Example, Basmati Rice)</P>
<P>DNA-Based Bar-Coded Signature of Fishes</P>
<P>Bioinformatics Tool and Protocol for SNP/STR Signatures</P>
<P>FastPCR</P>
<P>Suggested Readings</P>
<P>Important Links</P>
<P>Appendices</P>
<P>Glossary</P>
<P>Index</P>