Laboratory Protocols in Applied Life Sciences

1. Auflage 2014
ISBN: 978-1-4665-5314-9
Verlag: Taylor & Francis Inc

Buch, Englisch, 1826 Seiten, Format (B × H): 179 mm x 261 mm, Gewicht: 2606 g

As applied life science progresses, becoming fully integrated into the biological, chemical, and engineering sciences, there is a growing need for expanding life sciences research techniques. Anticipating the demands of various life science disciplines, Laboratory Protocols in Applied Life Sciences explores this development.

This book covers a wide spectrum of areas in the interdisciplinary fields of life sciences, pharmacy, medical and paramedical sciences, and biotechnology. It examines the principles, concepts, and every aspect of applicable techniques in these areas. Covering elementary concepts to advanced research techniques, the text analyzes data through experimentation and explains the theory behind each exercise. It presents each experiment with an introduction to the topic, concise objectives, and a list of necessary materials and reagents, and introduces step-by-step, readily feasible laboratory protocols.

Focusing on the chemical characteristics of enzymes, metabolic processes, product and raw materials, and on the basic mechanisms and analytical techniques involved in life science technological transformations, this text provides information on the biological characteristics of living cells of different origin and the development of new life forms by genetic engineering techniques. It also examines product development using biological systems, including pharmaceutical, food, and beverage industries.

Laboratory Protocols in Applied Life Sciences presents a nonmathematical account of the underlying principles of a variety of experimental techniques in disciplines, including:

Biotechnology
Analytical biochemistry
Clinical biochemistry
Biophysics
Molecular biology
Genetic engineering
Bioprocess technology
Industrial processes
Animal
Plant
Microbial biology
Computational biology
Biosensors
Each chapter is self-contained and written in a style that helps students progress from basic to advanced techniques, and eventually design and execute their own experiments in a given field of biology.

Zielgruppe

Graduate students and practitioners in a wide range of life sciences, as well as scientists and researchers in applied life sciences.

Autoren/Hrsg.

Bisen, Prakash Singh

Fachgebiete

• Medizin | Veterinärmedizin Medizin | Public Health | Pharmazie | Zahnmedizin Medizin, Gesundheitswesen Labormedizin
• Technische Wissenschaften Sonstige Technologien | Angewandte Technik Medizintechnik, Biomedizintechnik
• Naturwissenschaften Biowissenschaften Biowissenschaften
• Medizin | Veterinärmedizin Medizin | Public Health | Pharmazie | Zahnmedizin Medizin, Gesundheitswesen Medizintechnik, Biomedizintechnik, Medizinische Werkstoffe

Inhaltsverzeichnis

Microscopy
Introduction
Light Microscopy
Atomic Force Microscopy
Bright Field Microscopy
Dark Ground Microscopy
Phase Contrast Microscopy
Confocal Scanning Laser Microscopy
Differential Interference Contrast Microscopy
Polarization Microscopy
Fluorescence Microscopy
Experiments on Microscopy
Electron Microscopy
Sample Preparation for Electron Microscopy
Dark Field Electron Microscopy
Crewe Microscope
Backscatter Scanning Microscope
Experiment on Scanning Electron Microscopy of Cyanobacteria
Transmission Electron Microscopy of Phages (Bacteriophage and Cyanophage)
Suggested Readings
Important Links
Microtome
Introduction
Sectioning
Traditional Histology
Ultramicrotome
Cryostat
Incubation
Electron Microscopy Technique
Botanical Microtomy
Processing of Tissue
Suggested Readings
Important Links
pH and Buffers
Introduction
Acids and Bases
Hydrogen Ion Concentration and pH
Measurement of pH
Titration Curves
Buffer Solutions
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?
How the pH Changed on Adding 1 mL of 0.1 mol/L HCl in the Previous Mixture?
Buffers Used in Biochemical Experiments
pH and Life
Care and Use of the pH Meter
Titration Curves
Determination of pH Using Indicators
Titration of a Mixture of a Strong and a Weak Acid
Titration of a Strong Acid with a Strong Base
Titration of a Weak Acid with a Strong Base
Determination of pKa
pKa Values of a Dicarboxylic Acid
Acetate Buffers
Titration Curves of Amino Acids
Determination of pH of the Given Water/Soil Sample
Determination of pH of Fruit Juice
Measurement of pH of Soil Sample
Appendices
Suggested Readings
Important Links
Spectrophotometer
Introduction
Beer–Lambert Law
Limitations of the Beer–Lambert Law
Measurement of Extinction
Colorimeter
Spectrophotometry
Infrared Spectroscopy
Fluorescence Spectroscopy
Sensitivity of Fluorescence Assays
Fluorescence Quenching
Measurement of a-Naphthyl Phosphatase Activity
Suggested Readings
Important Links
Centrifugation
Introduction
Centrifuges
Ultracentrifugation
Suspending Medium
Rotors
Fractionation of Rat Liver
Estimation of DNA, RNA, and Protein in the Isolated Cell Fractions
Enzyme Distribution in the Cell
Glucose-6-Phosphatase
Glutamate Dehydrogenase
Acid Phosphatase
Mitochondrial Swelling
Determination of Lysosomal Integrity
Effect of Detergents on the Stability of the Lysosomal Membrane
Density Gradient Centrifugation
Fractionation of Pig Brain by Density Gradient Centrifugation
Distribution of Enzymes in the Fractions Obtained from Pig Brain
Isolation of Chloroplasts
Isolation of Chloroplast DNA
Isolation of Mitochondria and Assay of a Marker Enzyme
Isolation of Genomic DNA
Isolation of Chloroplasts from Spinach Leaves by Differential Centrifugation
Isolation of Plasmid DNA by Centrifugation
Suggested Readings
Important Links
Radiation Biology
Introduction
Radioisotopes
Sample Preparation
Autoradiography
Distribution of 14CO2 in Different Plant Parts
Photosynthetic Reduction of 14CO2 to Primary Metabolic Product
Incorporation of Labeled Acetate into Leucoplastic Fatty Acids of Developing Brassica Seeds
Suggested Readings
Important Links
Chromatography
Chromatography
Paper Chromatography
Ascending Chromatography
Descending Chromatography
Separation and Identification of Amino Acids by Descending Paper Chromatography
Separation and Identification of Amino Acids by Ascending Paper Chromatography
Separation and Identification of Amino Acids in a Given Mixture by 2 D Paper Chromatography
Thin Layer Chromatography
Thin-Layer Gel Chromatography
Separation and Identification of Sugars by Adsorption Thin Layer Chromatography
Identification of Lipids in a Given Sample by Thin Layer Chromatography
Identification of Sugars in Fruit Juices Using Thin Layer Chromatography
Column Chromatography
Gradient Preparation
Adsorption Chromatography
Separation of Pigments from Leaves or Flowers by Adsorption Column Chromatography
Ion-Exchange Chromatography
Separation of Amino Acids by Ion-Exchange Chromatography
Separation of Proteins from Human Serum by Ion-Exchange Chromatography
Gel Filtration/Size-Exclusion Column Chromatography
Separation of Blue Dextran and Cobalt Chloride on Sephadex G-25
Desalting of Protein Sample by Gel Filtration
Determination of Molecular Weight of a Given Protein by Gel Filtration
Concentration of Dilute Protein Solutions Using Sephadex G-25
Affinity Chromatography
Hydrophobic Interaction Chromatography or Affinity Elution
Purification by Affinity Chromatography
Preparation of an Affinity Column
Preparation of Rat Brain Acetylcholinesterase
Acetylcholinesterase Assay
Purification of Acetylcholinesterase by Affinity Chromatography
MALDI-TOF
Nuclear Magnetic Resonance Spectroscopy
Maintenance of Enzyme Activity
Addition of Chaotropes
Role of Polyols/Hydrophilic Polymers in Stabilizing Structures
Stabilization by Side-Chain Modification
Criteria of Purity
Biuret
Folin (Lowry’s Method)
Bradford
Suggested Readings
Important Links
Separation Technology
Electrophoresis
Principle
Types of Electrophoresis
Separation of Amino Acids by Paper Electrophoresis
Cellulose Acetate Strip Electrophoresis
Gel Electrophoresis
Polyacrylamide Gel Electrophoresis
Polyacrylamide Disc Gel Electrophoresis
SDS Gel Electrophoresis
Slab Gel Electrophoresis
Native Disc Gel Electrophoresis of Proteins
SDS-Polyacrylamide Slab Gel Electrophoresis of Proteins Under Reducing Conditions (SDS-PAGE)
Slab Gel Gradient Electrophoresis of Protein and Multiple Molecular Forms of Acetylcholinesterase
Protein (Western) Blotting
Isoelectric Focusing
Agarose Gel Electrophoresis
Determination of Molecular Weight of Plasmid DNA by Agarose Gel Electrophoresis
Electrophoretic Evaluation of Quality of High Molecular Weight DNA Isolated from Plant Material
Large-Scale Preparative Electrophoresis
Suggested Readings
Important Links
Histochemical Techniques
Introduction
Protein
Acidic Compounds
Carbohydrates–Polysaccharides
Lipids
Minor Compounds
Autoradiography
Enzyme Localization in Embedded Material
Suggested Readings
Important Links
Cytogenetics
Introduction
Study Different Stages in Root Tip Cells of Plant
Preparation of

Über Autor(innen)

Prakash S. Bisen received his PhD in 1972 and his DSc in 1981. He is currently pursuing his research in the field of medical biotechnology as emeritus scientist at Defense Research Development Establishment, Defense Research Development Organization, Ministry of Defense, Government of India, Gwalior, India. He has also been elected fellow of the National Academy of Sciences, India, for his enormous contributions to the field of biology. Bisen is the external scientific advisor to a Barcelona-based multinational company, Biokit, which is involved in the manufacturing of diagnostic kits. He has to his credit 200 publications in indexed international journals of repute, six international patents, as well as six international books on contemporary issues in biological sciences.