《EXPERIMENTAL MANUAL IN MEDICAL BIOCHEMISTRY》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 modern biochemical laboratory methodologies and techniques, develop the ability to perform experiments successfully and independently, properly interpret the data and write scientific laboratory reports.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. 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. All of the experiments have been performed many times in our own laboratory. We hope that they will provide a reliable training of the most important and commonly used techniques of contemporary biochemistry. We express our appreciation to our colleagues in the Department of Biochemistry and Molecular Biology, Wuhan University School of Medicine, who have contributed ideas, techniques and experiences to the manual. We are really grateful to all the experts, especially those from other universities, who made big contribution to the compiling of various chapters.

Progress in life science, including medical science, is mainly made by observation and experiment, especially biochemistry and molecular biology experiment. So, learning the basic biochemical techniques and methods is essential to medical students and the students of other related disciplines.
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 modern 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. All of the experiments have been performed many times in our own laboratory. We hope that they will provide a reliable training of the most important and commonly used techniques of contemporary biochemistry.
We express our appreciation to our colleagues in the Department of Biochemistry and Molecular Biology, Wuhan University School of Medicine, who have contributed ideas, techniques and experiences to the manual. We are really grateful to all the experts, especially those from other universities, who made big contribution to the compiling of various chapters.
This manual is not a complete version. It should be continually modified and updated. We would be very grateful if the users of this manual could bring good feedbacks for the development of this manual.

Chapter Ⅰ Spectrophotometry
1.1 Basic Concepts and Principle of SpectrophOtometry
1.2 Measu rement 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 Electropho retic 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 Com monly Used Chromatog raphiC 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 Enzvmarie 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 Pu rification
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.Transferase Fusion Protein
Chapter Ⅵ Isolation.Purification and Identification of Nucleic 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.Phenolor NaI Method
EXP.16 SDS.Alkaline Lysis of Plasmid DNA
EXP.17 Isolation of Total RNA by TRIzol Reagent
……
Chapter Ⅶ Genetic Engineering
English-Chinese Index