Practical Biotechnology : Methods and Protocols

The book helps undergraduate, postgraduate and research students to perform basic experiments in biotechnology. The laboratory protocols are simple to understand by students from different scientific backgrounds. Each laboratory exercise contains an introductory unit, protocol and easy-to-follow instructions for reagent preparation. The methods and protocols given here aim to make students ready for independent research in biotechnology laboratories. The protocols include * DNA Isolation * Molecular Biology Methods * RNA Isolation * Useful Information for Molecular Biology Methods * Working with DNA * Preparation of Solutions

Contents:

DNA ISOLATION - Isolation of Plasmid DNA / Isolation of Bacterial Genomic DNA / Isolation of Yeast Genomic DNA / Isolation of Fungal Genomic DNA / Isolation of Genomic DNA from Blood / Isolation of DNA from Animal Cells / Isolation of Genomic DNA from Eukaryotic Tissues / Isolation of Plant DNA using CTAB Extraction Method / Isolation of Chloroplast DNA / Mitochondrial DNA Isolation / Phenol Chloroform Extraction of DNA / Ethanol Precipitation of DNA / RNA ISOLATION - Isolation of Total RNA from Bacterial Cells / Isolation of Total RNA from Plant Tissues / Hot Phenol Isolation of RNA from Plant Tissues / Acid Phenol Extraction of RNA / Messenger RNA Isolation or Poly (A) RNA Isolation / WORKING WITH DNA - Storage / Purification / Concentration / Spectrophotometric Determination of Nucleic Acid Purity and Concentration / Fluorescent Quantification of DNA / Quantification of DNA using Diphenylamine (DPA) Assay / MOLECULAR BIOLOGY METHODS - Restriction Enzyme Digestion of DNA / DNA Ligation / Agarose Gel Electrophoresis of DNA / Elution of DNA Fragments from Agarose / Phenol Purification of DNA from Low Melting Agarose / Southern Blotting / Agarose Gel Electrophoresis of RNA / Northern Blotting / Cloning / Polymerase Chain Reaction (PCR)(In Vitro Amplification of DNA) / SDS Polyacrylamide Gel Electrophoresis / Western Blotting (Immunoblotting) / Iso-electric Focusing (IEF) of Proteins / 2D Gel Electrophoresis (2D PAGE) / Trypsin Digestion of Protein Gel / Protein Dialysis / Enzyme (Esterase) Gel Electrophoresis / SOME USEFUL INFORMATION FOR METHODS IN MOLECULAR BIOLOGY - Antibiotic Concentration in Media / E. coli Growth Curve / Storage of Bacterial Strains in Stab Agar / Storage of Bacterial Strains in Glycerol Solution / Decontamination of Ethidium Bromide (EtBr) Solutions / Preparation of Solutions / Glassware and Plasticware / Disposal of Buffers and Chemicals / Autoclave Operating Procedures / Safety Procedures / PREPARATION OF SOLUTIONS / References

Dr S Janarthanan is Senior Lecturer, Department of Zoology, Thiagarajar College, Madurai. He served as a Research Scientist for several years at Entomology Research Institute (ERI), Loyola College, Chennai (Madras). He has conducted and coordinated national and regional level training programmes in Techniques in Biotechnology sponsored by the Department of Biotechnology, Government of India, as well as ERI, Loyola College. Dr S Vincent is Reader, Department of Advanced Zoology and Biotechnology, Loyola College, Chennai. He has published several research papers in national and international journals and has co-authored the book Applied Genetics. He is a Steering Committee Member in the Ministry of Environment and Forests, Government of India.