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