The first edition of this manual presents shortened versions of the basic biochemistry methods designed for use at the lab bench in a biochemistry and molecular biology laboratory.It is primarily intended for undergraduate medical students, and hopefully it will be a useful reference book for graduate students and teaching staff. The objectives for the course are for students to learn modem biochemical laboratory methodologies and techniques, develop the ability to perform experiments successfully and independently, properly interpret the data and write scientific laboratory reports. Students who learn biochemistry as a part of medical science should be able to understand the concepts and methods of not only the conventional clinical laboratory tests but also the recently developed tests. For this purpose, we have made this manual with incorporation of some clinical laboratory test kits.
Chapter I Spectrophotometry
1.1 Basic Concepts and Principle of Spectrophotometnj
1.2 Measurement of Light Absorption
1.3 Aplications of Spectrophotometry
EXP. 1 Quantitative Analysis of Protein
EXP. 2 Determination of Blood Glucose
Chapter II Electrophoresis
2.1 Basic Principles of Electrophoresis
2.2 Impact Factors of Electrophoresis
2.3 Detection of Components after Electrophoretic Separation
2.4 Special Electrophoretic Techniques and Applications
EXP. 3 Separation of Serum Proteins by Cellulose Acetate Membrane
Electrophoresis
EXP. 4 Separation of Serum Lipoproteins by Agarose Gel Electrophoresis
EXP. 5 Proteins Separation by SDS-Polyacrylamide Gel Electrophoresis
Chapter III Chromatography
3.1 Fundamental Principles of Chromatography
3.2 Commonly Used Chromatographic Techniques
EXP. 6 Detecting Transamination of Amino Acids by Paper Chromatography
EXP. 7 Separation of Mixed Amino Acids by Cation Exchange
Chromatography
EXP. 8 Separation of Hemoglobin and Dinitrophenyl (DNP)-glutamate by
Gel Filtration Chromatography
Chapter IV Enzyme Analysis
4.1 The Activity of Enzyme
4.2 Enzyme kinetics (Factors affecting reaction velocity)
4.3 Enzymatic analysis
EXP. 9 Assay the Activity of Alanine Aminotransferase (ALT) in Serum
( Mohun's Method)
EXP. 10 Lactate Dehydrogenase (LDH) Analysis
EXP. 11 Effect of Substrate Concentration on Enzyme Activity-Determining
Km Value for Alkaline Phosphatase
EXP. 12 Coupled enzymatic reaction assay (EnzymaticEndpoint Method)-
Determination of Total Cholesterol, Triglycerides and
Glucose in Serum
Chapter V Isolation and Purification of Protein
5.1 Methods and Basic Principles of Protein Purification
5.2 Procedure of Protein Preparation
5.3 Crystallization, Concentration, Drying and Storage of Protein
5.4 Identification and Analysis of Protein
EXP. 13 Isolation and Identification of Serum IgG
EXP. 14 Expression, Purification and Identification of Glutathione S-Transferas
Fusion Protein
Chapter VI Isolation, Purification and Identification of Nucle Acids
6.1 Isolation and Purification of Nucleic Acids
6.2 Identification and Analysis of Nucleic Acids
EXP. 15 Isolation of Eukaryotic Genomic DNA by Proteinase K-Phenol
or NaI Method
EXP. 16 SDS-Alkaline Lysis of Plasmid DNA
EXP. 17 Isolation of Total RNA by TRIzol Reagent
EXP. 18 Identification of DNA by UV-Spectrophotometry and
Gel Electrophoresis
EXP. 19 Identification of RNA by UV-Spectrophotometry and Formaldehyde Denaturating Agarose Gel Electrophoresis
EXP. 20 Polymerase Chain Reaction (PCR) and Reverse Transcription-PCR
Chapter VII Genetic Engineering
7.1 The Process of Gene Cloning
7.2 Enzymes for Gene Cloning
7.3 Vectors for Gene Cloning
7.4 Expression of Foreign Genes and Purification of Recombinant Proteins
EXP. 21 DNA Cloning
EXP. 22 Expression of Exogenous Gene in E. coli and Purification of the Expressional Product
English-Chinese Index