Main description:

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.

Contents:

Section 1:

(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.