Basic Techniques in Biochemistry, Microbiology and Molecular Biology
Principles and Techniques
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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.
(A) Equipmentsi. BOD incubatorsii. Laminar flowiii. Biosafety Cabinetsiv. Aseptic hoodv. Autoclavevi. Hot air sterilizervii. Deep freezerviii. Refrigeratorix. Microscopyx. Colony counterxi. Nebular chamberxii. Turbidometerxiii. Colorimeterxiv. Spectrophotometerxv. PCRxvi. ELISA readerxvii. Sonicator
(B) Practical Molecular Biology 1. Enzyme assay: qualitative and quantitative (protease, amylase, penicillinase, pectinolytic enzyme, chitinase, keratinase)2. Antimicrobial sensitivity assay 3. Extraction of DNA and RNA: Plant cells, Animal cells and Microorganism4. Estimation of DNA by diphenylamine reaction5. Estimation of RNA using orcinol method6. Determination of melting temperature of DNA7. Quantitative estimation of DNA and RNA8. Purification and bioassay of interleukin 1 and 2 (IL 1 and IL 2)9. Quantitative determination of TNF alpha and Interleukin 6 in the given sample10. Quantitative determination of amount of mono and oligo nucleosomes in the cytosol of cell extract sample after an induced cell death.(C) Practical Microbiology
1. Sterilization of glassware, preparation of media and its sterilization2. Sub culturing of bacteria, fungi and actinomycetes.3. Preservation of microorganisms: stabs, slants, lyophilization, cryopreservation 4. Staining methods: Simple, negative, Gram staining, Acid-fast staining, cell wall, capsule, flagella, nuclear material, cytoplasm, filamentous fungi, mycoplasma5. Isolation and purification of microorganisms: streaking, pour plate, spread plate, serial dilution, Warcup method, baiting technique, enrichment culture6. Isolation of coprophilous fungi 7. Isolation of microorganisms from air, water 8. Isolation of anaerobic microorganisms 9. Isolation of extremophiles: thermophiles and psychrophiles10. Microbiological assay of antibiotics by cup plate method11. Motility testing: hanging drop method and stab12. Sterility testing of pharmaceuticals13. Physical, Chemical and Bacteriological analysis of water14. Biochemical identification of microorgamism: IMViC, oxidative fermentation, carbohydrate and nitrogen source utilization15. Immobilization of microorganisms and enzymes
Section 2:1. Qualitative systematic analysis of carbohydrates (Glucose, Fructose, Lactose, Maltose, Sucrose and starch).2. Protein analysis in food.3. Identification tests for Proteins (Albumin and Casein).4. Quantitative analysis of reducing sugars by 3, 5-Dinitrosalicylic acid (DNSA method). 5. Quantitative analysis of Proteins by various methods including Biuret.6. Qualitative analysis of urine for abnormal constituents.7. Determination of blood creatinine.8. Determination of blood sugar.9. To perform oral glucose tolerance test.10. Determination of serum total cholesterol.11. Preparation of buffer solution and measurement of pH.12. Study of enzymatic hydrolysis of starch.13. Determination of Salivary amylase activity.14. Study the effect of Temperature on Salivary amylase activity.15. Study the effect of substrate concentration on salivary amylase activity.16. Analysis of Butter.17. Analysis of Milk.18. TLC and paper chromatography of amino acid and carbohydrates.19. Fat characterization20. Analysis of bread.
RICHA JAIN is a Senior Scientist, working in the Centre for Scientific Research and Development, People's University, Bhopal, India, possesses more than 14 years of research experience in the area of molecular biology, animal cell culture, microbiology, bioremediation, and industrial and therapeutic enzymes. Presently, she is working on two projects: (a) therapeutic enzymes arginases (anticancer) and (b) bioremediation of plastic waste. She has published 21 research papers with 81 citations in research journals of high repute. Among several feathers on her cap includes Best Paper Award for the presentation of her paper on "Bioinformatics: Importance of Proteomics and Genomics" during the National Conference on "New Frontiers of Sciences" National Science Day Celebrations 2003, Dr. Hari Singh Gour University, Sagar, 28 February 2003; All India Young Scientist Award for the presentation of her paper "Production of Proteins and Amino Acids Using Poultry Feathers from a Protease Producing Strain of Streptomyces sp. CFS 1068" during Bhartiya Vigyan Sammelan 2007 in Silver Jubilee of All India Young Scientist Conference, M.P. Council of Science and Technology, Bhopal, 23-25 November 2007; 1st Prize for her paper presentation on "Fluconazole-Loaded Cubosome: An Excellent Nanoparticle for Treatment of Cutaneous Candidiasis" during the National Seminar on Recent Advancement in Drug Delivery Techniques and Drug Designing, School of Pharmacy Research, Peoples University, Bhopal, 26-27 September 2014; and Appreciation for Oral Presentation in the International Conference on Translational Medicine in the Twenty-First-Century "Stem Cell Transplantation: Current Status," Barkatullah University, Bhopal, 11-14 April 2015. She has also presented paper entitled "Study of Anti-adenocarcinoma Activity of Arginase by Streptomyces Species." She is also a Member of different societies like the Association of Microbiologists of India, Indian Science Congress Association, Institutional Biosafety Committee, etc. She has actively participated in more than 30 scientific workshops and conferences and has also delivered invited guest lectures in workshops and conferences along with providing hands-on training in eight workshops.
SOURABH JAIN is an Associate Professor at Sagar Institute of Pharmaceutical Sciences, Sagar, India. He has completed his PhD from Shobhit University, Meerut, UP, India; MPharm from Smriti College of Pharmaceutical Education, Indore, MP, India; and BPharm from B.R. Nahata College of Pharmacy, Mandsaur, MP, India. He is having 8 years of experience in academic and research. He has published 24 papers in national and international journals and conferences. His key research interests include standardization of Ayurvedic and polyherbal formulation, antidiabetic screening and phytochemical isolation of medical plant, and biochemical testing.