A laboratory Text book of Biochemistry, Molecular Biology and Microbiology is intended to prepare the undergraduate, postgraduate and research students to perform basic experiments on various aspects of bioscience and biotechnology. Moreover, in the Semester system of teaching it is necessary to explore experiments which are not lengthy and easily completed within contact hours. Initially the book deals with dilutions, pH, buffers, units of measurements and calculations. This is followed by lab safety rules which is very important for any student working with chemicals for their and safety of others. This book emphasizes on principles, reagent preparations and procedures related to experiments, which will be handy for students from different scientific backgrounds. A number of methods are available in the literature for quantification of various molecules. This book does not present all the available methods but based on experience it contains commonly used methods, which students should know. The methods have been written in a manner for direct practical use in the laboratory.
This work has originated as a result of numerous requests from my students for eased out and explanatory methods pertaining to biochemistry, biotechnology, microbiology and others. The section on testing of adulterants is of much use for common mass because most of the food products we eat are adulterated. The approach is rather simple with the use of very easily available chemicals and the tests can be performed even in house. It is hoped that the reliable assays presented in this manual will help the students and research scholars to get to basics of experiments and various aspects associated with it.
Inhaltsverzeichnis (Table of Contents)
- WATER, PH AND BUFFERS.
- PREPARATION OF ACETATE BUFFER
- TESTING OF BUFFER CAPACITY
- DILUTIONS, UNITS FOR MEASURING CONCENTRATION OF SOLUTIONS AND SPECTROPHOTOMETRIC CALCULATIONS
- LAB SAFETY RULES.
- CARBOHYDRATES..
- DETERMINATION OF REDUCING SUGARS CONTENT (NELSON-SOMOGYI METHOD).
- DETERMINATION OF SUGARS CONTENT (DNSA METHOD).
- DETERMINATION OF TOTAL CARBOHYDRATES (ANTHRONE METHOD)
- DETERMINATION OF TOTAL CARBOHYDRATES (PHENOL SULFURIC ACID METHOD)..
- DETERMINATION OF GLUCOSE CONTENT (GLUCOSE OXIDASE METHOD)...
- DETERMINATION OF STARCH CONTENT.
- DETERMINATION OF CELLULOSE CONTENT
- DETERMINATION OF FRUCTOSE CONTENT.
- DETERMINATION OF INULIN CONTENT
- LIPIDS...........
- QUALITATIVE TESTS FOR LIPIDS.
- QUALITATIVE TESTS FOR CHOLESTEROL
- DETERMINATION OF CHOLESTEROL CONTENT (ZAK'S METHOD) ......
- DETERMINATION OF OIL/LIPID CONTENT IN OILSEEDS
- DETERMINATION OF VOLATILE ACIDS CONTENT.
- DETERMINATION OF ACID VALUE AND % FREE FATTY ACIDS ....
- DETERMINATION OF SAPONIFICATION NUMBER OF OIL/FAT.
- DETERMINATIONOF IODINE NUMBEROF OIL/FAT.
- DETERMINATION OF PEROXIDE VALUE...
- SEPARATION OF PLANT PIGMENTS USING THIN LAYER CHROMATOGRAPHY
- PROTEINS.
- QUALITATIVE TEST FOR AMINO ACIDS AND PROTEINS..
- PRECIPITATION OF PROTEINS (A GENERAL APPROACH)...
- DETERMINATION OF MAX FOR A GIVEN PROTEIN SAMPLE.
- DETERMINATION OF PROTEIN CONTENT (LOWRY'S METHOD).
- PRECIPITATION AND DETERMINATION OF PROTEIN CONTENT (TCA METHOD)..
- DETERMINATION OF PROTEIN CONTENT (BIURET METHOD).
- DETERMINATION OF PROTEIN CONTENT (BRADFORD METHOD).
- DETERMINATION OF PROLINE CONTENT IN PLANTS .
- ISOLATION AND PURIFICATION OF CASEIN FROM MILK
- DETERMINATION OF TOTAL FREE AMINO ACIDS
- DETERMINATION OF TRYPTOPHAN CONTENT.
- DETERMINATION OF METHIONINE CONTENT
- DETERMINATION OF LYSINE CONTENT
- SEPARATION OF AMINO ACIDS BY CIRCULAR PAPER CHROMATOGRAPHY
- SEPARATION OF AMINO ACIDS USING PAPER CHROMATOGRAPHY...
- SDS-PAGE.
- PREPARATION OF STANDARD CURVE OF PARA NITRO PHENOL (PNP)
- ENZYMATIC ASSAY OF ACID PHOSPHATASE.
- DETERMINATION OF THE TIME LINEARITY OF AN ENZYMATIC REACTION
- DETERMINATION OF THE PROTEIN LINEARITY OF AN ENZYMATIC REACTION
- TO DETERMINE THE PROPERTIES OF (KM AND VMAX VALUES) OF AN ENZYME..
- NITRATE REDUCTASE ASSAY
- AMYLASE ASSAY
- PHENYL AMMONIA LYASE ASSAY
- NUCLEIC ACIDS
- ISOLATION OF GENOMIC DNA
- SOLATION OF PLASMID..
- ISOLATION OF PLANT GENOMIC DNA..
- DETERMINATION OF DNA CONTENT (DIPHENYL AMINE TEST) .............
- DETERMINATION OF CONCENTRATION AND PURITY OF THE DNA SAMPLE..
- ELECTROPHORESIS
- RESTRICTION DIGESTION OF DNA.
- SOUTHERN BLOTTING
- ISOLATION OF TOTAL RNA FROM PLANT TISSUE
- ISOLATION OF RNA FROM YEAST.
- DETERMINATION OF RNA CONTENT (ORCINOL REAGENT METHOD).
- MICROBIOLOGY .....
- PREPARATION OF MEDIA
- PREPARATION OF SLANTS, STABS AND POURING ON PETRIPLATES....
- GRAM STAINING OF BACTERIA
- STAINING OF BACTERIAL SPORES.
- STREAK PLATE TECHNIQUE
- ASSESSMENT OF AIR MICROFLORA.
- ENUMERATION OF BACTERIA IN SOIL............
- PREPARATION OF Escherichia coli COMPETENT CELLS AND ITS TRANSFORMATION
- STUDY OF DIFFERENT GROWTH PHASES OF BACTERIAL POPULATION AND PLOT A BACTERIAL GROWTH CURVE.
- ANTIMICROBIAL SUSCEPTIBILITY TESTING (DISC DIFFUSION METHOD)
Zielsetzung und Themenschwerpunkte (Objectives and Key Themes)
This laboratory textbook aims to equip undergraduate, postgraduate, and research students with the necessary skills to perform fundamental experiments in bioscience and biotechnology. It focuses on providing practical, concise methods that can be completed within typical lab hours. The text covers a wide range of topics, including: * **Fundamentals of Laboratory Techniques**: The book begins with essential techniques such as dilutions, pH measurements, buffer preparation, and lab safety protocols. * **Biomolecules and their Analysis**: The text then delves into the analysis of various biomolecules, including carbohydrates, lipids, proteins, and nucleic acids. This includes qualitative and quantitative assays for these molecules, along with techniques like chromatography and electrophoresis. * **Microbiology Fundamentals**: The book concludes with a comprehensive introduction to basic microbiology techniques, covering media preparation, bacterial staining, and various microbiological assays.Zusammenfassung der Kapitel (Chapter Summaries)
The book begins with chapters on fundamental lab techniques, covering water, pH, buffers, dilutions, units of measurement, and spectrophotometric calculations. It then emphasizes lab safety protocols, essential for students working with chemicals. The following chapters delve into the analysis of biomolecules. The carbohydrate section explores various methods for determining the content of different sugars, including reducing sugars, total carbohydrates, and specific sugars like glucose, starch, cellulose, fructose, and inulin. The lipids chapter covers qualitative tests for lipids and cholesterol, and methods for determining cholesterol content, oil/lipid content in oilseeds, volatile acids content, acid value, saponification number, iodine number, and peroxide value. It also includes a section on the separation of plant pigments using thin-layer chromatography. The proteins chapter explores qualitative tests for amino acids and proteins, precipitation methods, determination of protein content using various methods (Lowry, TCA, Biuret, Bradford), and analyses of specific amino acids like proline, tryptophan, methionine, and lysine. It also covers techniques for separating amino acids using chromatography and electrophoresis. The nucleic acids chapter covers the isolation of DNA and RNA, determination of their content, and techniques like electrophoresis, restriction digestion, and Southern blotting. The final chapters introduce basic microbiology techniques, including media preparation, staining methods, culturing techniques, assessment of air microflora, bacterial enumeration in soil, and antimicrobial susceptibility testing.Schlüsselwörter (Keywords)
This textbook focuses on fundamental laboratory techniques, biomolecule analysis, and basic microbiology methods. Key concepts include: dilutions, pH, buffers, spectrophotometric calculations, lab safety, carbohydrates, lipids, proteins, nucleic acids, chromatography, electrophoresis, media preparation, bacterial staining, and antimicrobial susceptibility testing.- Quote paper
- Sharad Vats (Author), 2014, A laboratory Text book of Biochemistry, Molecular Biology and Microbiology, Munich, GRIN Verlag, https://www.grin.com/document/284414
