TY - BOOK AU - Thatoi, Hrudayanath AU - Dash, Supriya AU - Das, Swagat Kumar TI - Practical Boitechnology : Principles and Protocols SN - 9789389633931 U1 - 660.6 PY - 2019/// CY - New Delhi PB - Dreamtech press/ I.K. International Pvt. Ltd. KW - Biotechnology N1 - Contents Preface Acknowledgements 1. General Laboratory Principles 1.1 Safety Rules 1.2 Cleaning Glassware and Plasticware 1.3 Basic Laboratory Techniques Review Questions 2. Biochemistry 2.1 Determination of pKa Value 2.2 Validation of Beer-Lambert Law 2.3 Determination of Absorption Maxima 2.4 Calibration Curve of Starch 2.5 Carbohydrates 2.6 Quantitative Analysis of Carbohydrates 2.7 Estimation of Proteins 2.8 Estimation of Amino Acids 2.9 Estimation of Lipid 2.10 Estimation of DNA 2.11 Estimation of RNA 2.12 Estimation of Ascorbic Acid in Plant Materials 2.13 Estimation of Thiamine 2.14 Quantitative Estimation of Phosphate Solubilisation in Culture Medium Review Questions 3. Molecular Biology-I (Genomic Technology) 3.1 Isolation of Genomic DNA 3.2 Isolation of Plasmid DNA 3.3 Isolation of RNA 3.4 DNA Quantification and Quality Analysis 3.5 Agarose Gel Electrophoresis for DNA Quantification and Quality Analysis 3.6 Restriction Digestion 3.7 PCR Amplification 3.8 DNA Fingerprinting Technique 3.9 DNA Sequencing 3.10 Blotting Technique 3.11 Cloning Review Questions 4. Molecular Biology-II (Proteomics Techniques) 4.1 Protein Precipitation by Salting Out Method 4.2 Dialysis 4.3 Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis (SDS-PAGE) of Proteins 4.4 Native-PAGE 4.5 2-D Electrophoresis Review Questions 5. Immunological Techniques 5.1 Immunodiffusion Techniques 5.2 Immunoelectrophoresis 5.3 Rocket Immunoelectrophoresis 5.4 Counter Current Immunoelectrophoresis 5.5 Enzyme-Linked Immunosorbent Assay (ELISA) 5.6 Antibody Production 5.7 Assay of Non-Specific Immune Parameters Review Questions 6. Cytogenetics and Haematology 6.1 Study of Different Stages of Mitosis 6.2 Study of Different Stages of Meiosis 6.3 Karyotyping 6.4 Blood Grouping 6.5 Differential White Blood Cell Count (Differential Leukocyte Count) 6.6 Total Red Blood Cell Count 6.7 Estimation of Haemoglobin (Hb) Review Questions 7. Microbial Techniques 7.1 Cleaning Glassware 7.2 Sterilization 7.3 Preparation of Culture Media 7.4 Techniques for Isolation of Pure Cultures 7.5 Gram Staining 7.6 Acid-Fast Staining 7.7 Fungal Staining 7.8 Mycorrhizal Staining 7.9 Isolation of Cyanobacteria 7.10 Isolation of Rhizobium 7.11 Microbial Growth Curve 7.12 Isolation of Bacteriophage 7.13 Isolation of Tobacco Mosaic Virus (TMV) 7.14 Antibiotic Assay 7.15 Bacterial Sensitivity 7.16 Paddy Straw Mushroom Cultivation Review Questions 8. Fermentation and Enzyme Technology 8.1 Starter Culture Preparation for Bioethanol Production 8.2 Production of Ethanol 8.3 Ethanol Fermentation by Immobilization of Yeast 8.4 Estimation of Alcohol 8.5 Wine Production 8.6 Beer Production 8.7 Bioreactor Designing 8.8 Amylase Production 8.9 Enzymatic Assay Review Questions 9. Plant Tissue Culture Techniques 9.1 Preparation of Culture Media 9.2 Callus Culture 9.3 Direct Organogenesis: Shoot Tip Culture 9.4 Embryo Culture 9.5 Cell Suspension Culture 9.6 Anther Culture 9.7 Induction of Hairy Roots using Agrobacterium Rhizogenes 9.8 Agrobacterium Mediated Biotransformation 9.9 Artificial Seeds/Synthetic Seeds 9.10 Protoplast Culture 9.11 Isolation of Protoplast by Mechanical Method 9.12 Isolation of Protoplast by Enzymatic Method 9.13 Gene Transfer by Particle Bombardment 9.14 Gene Transfer by Microinjection Technique 9.15 Gene Transfer by Electroporation Technique 9.16 PEG-Mediated Gene Transfer Review Questions 10. Animal Cell Culture 10.1 Preparation of Media 10.2 Primary Cell Culture 10.3 Preparation of Established Cell Lines 10.4 Maintenance of Cell Lines 10.5 Preservation of Cell Lines Review Questions 11. Pharmaceutical Bioprocessing Techniques 11.1 Extraction Techniques used for Isolating Phytochemicals from Medicinal Plants 11.2 Liquid-Liquid Extraction Technique 11.3 Chromatography 11.4 Qualitative Phytochemical Assay 11.5 Estimation of Total Flavonoid Content 11.6 Estimation of Total Tannin Content 11.7 Cytotoxicity Study (MTT Assay) Review Questions 12. Analytical Procedures for Water Analysis 12.1 Dissolved Oxygen (DO) 12.2 Biochemical Oxygen Demand (BOD) 12.3 Chemical Oxygen Demand (COD) 12.4 Estimation of Phosphate (PO43–) 12.5 Estimation of Ammonia (NH3) 12.6 Estimation of SO42– (Sulphate) 12.7 Estimation of Nitrite 12.8 Estimation of Nitrate (NO3–) 12.9 Estimation of Chloride (Cl–) 12.10 Estimation of Total Hardness 12.11 Determination of Total Alkalinity (Richards, 1954) 12.12 Estimation of Total Dissolved Solid 12.13 Estimation of Hexavalent Chromium Review Questions 13. Biostatistics 13.1 Mean, Median, Mode, Standard Deviation 13.2 T-Test 13.3 Regression and Correlation 13.4 Analysis of Variance (ANOVA) Review Questions 14. Bioinformatics Techniques 14.1 Database Search 14.2 BLAST (Basic Local Alignment Search Tool) 14.3 FASTA 14.4 Multiple Sequence Alignment 14.5 Study of Human Genome Data 14.6 Homology Modelling Review Questions 15. Biophysical Techniques (Bioinstrumentation) 15.1 Laminar Airflow 15.2 Centrifuge 15.3 Lyophilizer 15.4 Microscopy 15.5 UV-VIS Spectrophotometer 15.6 Electrophoresis 15.7 Gel Documentation System 15.8 UV-Transilluminators 15.9 High Performance Liquid Chromatography (HPLC) 15.10 Thermal Cycler 15.11 Microplate Reader 15.12 Infrared Spectroscopy 15.13 Nuclear Magnetic Resonance (NMR) 15.14 Mass Spectroscopy 15.15 X-Ray Diffraction 15.16 Microarray Review Questions Index N2 - This book principally contains practical aspects of biotechnology, its theoretical background, working principles, and procedures meant for conducting experiments in the laboratories. The experiments in this book are spread in different chapters like Biochemistry, Microbiology, Immunology, Molecular biology, Plant and Animal Biotechnology, Environmental Technology, Industrial Biotechnology, Biostatistics and Bioinformatics. Besides, a chapter has been devoted towards discussion on safety guidelines, setting up of biotechnology laboratory, ethical issues concerning microbiology and genetic engineering work along with different reagent preparation for conducting laboratory experiments. Each chapter aims to describe both the theory and relevant practical details for a given technique, and to identify both the potential and limitations of the technique. This book is designed as an intensive introduction to the various tools of molecular biology. It introduces all the basic methods of molecular biology including cloning, PCR, Southern blotting, Northern blotting, Western blotting, DNA sequencing. This book is useful for final year undergraduate (especially project) students, post-graduate research students and research scientists and technicians who wish to understand and use new techniques in the field of Biotechnology ER -