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Basic Techniques in Biochemistry, Microbiology and Molecular Biology: Principles and Techniques (Springer Protocols Handbooks)

✍ Scribed by Aakanchha Jain, Richa Jain, Sourabh Jain


Publisher
Springer
Year
2020
Tongue
English
Leaves
264
Edition
1st ed. 2020
Category
Library

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✦ Synopsis


This book presents key methodologies, tools and databases for biochemistry, microbiology and molecular biology in simple and straightforward language. Covering all aspects related to experimental principles and procedures, the protocols included here are brief and clearly defined, and include essential precautions to be taken while conducting experiments. The book is divided into two major sections: one on constructing, working with, and standard operating procedures for laboratory instruments; and one on practical procedures used in molecular biology, microbiology and biochemical analysis experiments, which are described in full.

Each chapter describes both the basic theory and relevant practical details for a given experiment, and helps readers recognize both the experiment’s potential and limitations. Intended as an intensive introduction to the various tools used in molecular biology, the book covers all basic methods and equipment, including cloning, PCR, spectrophotometers, ELISA readers, sonicators, etc. As such, it offers a valuable asset for final year undergraduate (especially project) students, graduate research students, research scientists and technicians who wish to understand and employ new techniques in the field of biotechnology.

✦ Table of Contents


Preface
Contents
About the Authors
Part I: Instruments
Chapter 1: BOD (Biochemical Oxygen Demand or Biological Oxygen Demand) Incubator
Chapter 2: Laminar Air Flow/Biosafety Cabinets
Chapter 3: Aseptic Hood
Chapter 4: Autoclave
Chapter 5: Hot Air Oven
Chapter 6: Deep Freezer (-20 C) (Low-Temperature Cabinet)
Chapter 7: Refrigerator
Chapter 8: Compound Microscope
Chapter 9: Digital Colony Counter
Chapter 10: Digital Turbidity Meter
Chapter 11: Digital Nephelometer
Chapter 12: Digital PhotoColorimeter
Chapter 13: Digital UV/Visible Spectrophotometer
Chapter 14: Polymerase Chain Reaction
Chapter 15: ELISA Reader
Chapter 16: Sonicator
Part II: Molecular Biology
Chapter 17: Enzyme Assay: Qualitative and Quantitative
1 Qualitative Enzyme Assays
2 Quantitative Enzyme Assay
Chapter 18: Antimicrobial Sensitivity Assay
1 Procedure
2 Procedure
Chapter 19: Extraction of DNA from Plant Cells
1 From Onion
Procedures
2 From Spinach and Cauliflower
Chapter 20: Extraction of DNA from Animal Cells
1 From Blood [Using Phenol Extraction Method]
2 From Blood [Using Isopropanol Precipitation]
3 From Mouse Liver
4 From Pig Spleen
Chapter 21: Extraction of DNA and RNA from Microorganism
1 From Bacteria
2 From Yeast Cells
Chapter 22: Estimation of DNA by Diphenylamine Reaction
Chapter 23: Estimation of RNA Using Orcinol Method
Chapter 24: Determination of Melting Temperature of DNA
Chapter 25: Quantitative Estimation of DNA and RNA
Chapter 26: Purification and Bioassay of Interleukin-1 and Interleukin-2 (IL-1 and IL-2)
Chapter 27: Quantitative Determination of TNF Alpha and Interleukin-6
Part III: Microbiology
Chapter 28: Sterilization of Glassware; Preparation and Sterilization of Media
Chapter 29: Sub-culturing of Bacteria, Fungi and Actinomycetes
Chapter 30: Preservation of Microorganisms: Stabs, Slants, Lyophilization and Cryopreservation
Chapter 31: Staining Methods - Simple Staining, Negative Staining, Gram´s Staining and Acid-Fast Staining
Chapter 32: Isolation of Coprophilous Fungi (Moist Chamber Method)
Chapter 33: Isolation of Microorganisms from Air
Chapter 34: Motility Testing - Hanging Drop Method and Stab
Chapter 35: Sterility Testing of Pharmaceuticals
1 Method B: Direct Inoculation (Table 7)
Chapter 36: Physical, Chemical and Bacteriological Analysis of Water
Methods
Ultraviolet Spectrophotometric Method
Nitrate Electrode Method
Phenoldisulfonic Acid (PDA) Method
Chapter 37: To Perform Biochemical Identification of Microorganism: IMViC (Oxidative Fermentation and Carbohydrate Source Util...
Chapter 38: To Perform Biochemical Identification of Microorganism by Nitrogen Source Utilization or Urease Test
Chapter 39: Immobilization of Enzymes and Microorganisms
1 Enzyme Immobilization in Polyacrylamide Gel
2 Enzyme Immobilization in Alginate Gel
3 Enzyme Immobilization in Gelatin Gel
4 Cell Immobilization in Agarose
5 Immobilization of Microbial Cells in Gelatin
Part IV: Biochemistry
Chapter 40: Qualitative Systematic Analysis of Carbohydrates (Glucose, Fructose, Lactose, Maltose, Sucrose and Starch)
1 Molisch´s Test (α-Naphthol Reaction)
2 Benedict´s Test
3 Fehling´s Test
4 Iodine Test
5 Nylander´s Test (Bismuth Reduction Test)
6 Cole´s Test
7 Reduction of Methylene Blue Test
8 Barfoed´s Test: Reducing Monosaccharide
9 Bial´s Test for Pentose
10 Seliwanoff´s Test
11 Hydrolysis of Sucrose
12 Methylamine Test for Lactose
13 Osazone Test
Chapter 41: Protein Analysis in Food
1 Kjeldahl Method
2 Dumas (Nitrogen Combustion)
3 Infrared Spectroscopy
4 Colorimetric Methods
5 Dye-Binding Methods
5.1 Anionic Dye-Binding Method
5.2 Bradford Dye-Binding Method
6 Copper Ion-Based Methods
6.1 Biuret Method
6.2 Lowry Method
6.3 Bicinchoninic Acid Method
7 Ultraviolet Absorption at 280 nm
8 Peptide Measurement at 190-220 nm
Chapter 42: Identification Tests for Proteins (Casein and Albumin)
Chapter 43: Quantitative Analysis of Reducing Sugars by 3, 5-Dinitrosalicylic Acid (DNSA Method)
Chapter 44: Quantitative Analysis of Proteins by Various Methods Including Biuret
1 Other Methods
1.1 Bradford Method
1.2 Lowry Method
1.3 Protein Precipitation
1.3.1 Ammonium Sulfate Precipitation
Chapter 45: Qualitative Analysis of Urine for Abnormal Constituents
1 Physical Properties
2 Biochemical Characterization of Urine to Detect Abnormal Constituents
3 Benedict´s Test
4 Tauber´s Test for Detection of Aldopentoses in Urine Samples
5 Bial´s Test for Detection of Pentoses in Urine Samples
6 Seliwanoff´s Test for Detection of D-Fructose in Urine Samples
7 Mucic Acid Test for Analysis of Lactose or Galactose
8 Detection of Acetone Bodies Using Rothera´s Nitroprusside Test
9 Estimation of Acetoacetic Acid by Gerhardt´s Test
10 Analysis of Proteins in Urine Sample by Sulfosalicylic Acid Test
11 Analysis of Proteins in Urine Sample by Bence Jones Protein
12 Determination of Bile Pigments by Harrison Spot Test
13 Schlesinger´s Test for Urobilinogen
14 Detection of Bile Salts in Urine by Pettenkofer´s Test
15 Detection of Occult Blood in Urine
Chapter 46: Determination of Blood Creatinine
1 Serum Creatinine Test (Jaffe´s Test)
Chapter 47: Determination of Blood Sugar
1 Folin-Wu Method
2 Glucose Oxidase Method
Chapter 48: To Perform Oral Glucose Tolerance Test
Chapter 49: Determination of Serum Total Cholesterol
1 Enzymatic Colorimetric Method
2 Sackett´s Method
3 Zak´s Method
Chapter 50: Preparation of Buffer Solution and Measurement of pH
Chapter 51: Study of Enzymatic Hydrolysis of Starch
Chapter 52: Determination of Salivary Amylase Activity
Chapter 53: Study the Effect of Temperature on Salivary Amylase Activity
Chapter 54: Study the Effect of Substrate Concentration on Salivary Amylase Activity
Chapter 55: Analysis of Butter
Chapter 56: Analysis of Milk
Chapter 57: Thin Layer Chromatography of Carbohydrates
Chapter 58: Thin Layer Chromatography of Amino Acid
Chapter 59: Paper Chromatography of Amino Acid
Chapter 60: Paper Chromatography of Carbohydrates
Chapter 61: Fat Characterization
1 Methods for Characterization of Fats
1.1 Melting Point
1.2 Iodine Value
1.3 Saponification Value
1.4 Peroxide Value
2 Methods for Lipid Components
2.1 Fatty Acid Composition and Fatty Acid Methyl Esters (FAMEs)
2.2 Cholesterol and Phytosterols
Chapter 62: Analysis of Bread
Appendices
Appendix I: Composition of Different Types of Media
Appendix II: Buffers
Bibliography


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