生物化学与分子生物学实验教程

Part 1 Basic techniques of biochemistry and molecular biology

Chapter 1 Basic techniques of preparation of biomacromolecules

Section 1 Salting out

Section 2 Dialysis and ultrafiltration

Chapter 2 Spectrophotometry

Section 1 Basic principles

Section 2 The basic construction of spectrophotometers

Section 3 Some home-made spectrophotometers

Chapter 3 Chromatography

Section 1 Adsorption chromatography

Section 2 Partition chromatography

Section 3 Ion-exchange chromatography

Section 4 Gel filtration

Section 5 Affinity chromatography

Chapter 4 Electrophoretic techniques

Section 1 Basic principles

Section 2 Several types of commonly used electrophoresis

Chapter 5 Centrifugation techniques

Section 1 Basic principle

Section 2 General description of centrifugation

Chapter 6 Genetic engineering and analytieal technology of gene expression

Section 1 Genetic engineering

Section 2 Polymerase chain reaction

Section 3 Nucleic acid hybridization

Section 4 DNA sequencing technology

Part 2 Experiments of the biochemistry and molecular biology

Chapter 7 Analysis of purification and physicochemical properties of proteins

Experiment 1 Colorimetric methods for protein determination

Experiment 2 Isolation and purification of serum y-globulin

Experiment 3 Cellulose acetate membrane electrophoresis of serum proteins

Experiment 4 Determination of relative molecular mass of protein in SDS-PAGE

Chapter 8 Kinetics of enzyme-catalysed reactions

Experiment 1 The specificity of enzyme and effect of temperature， pH on the enzyme Activity

Experiment 2 Isolation of alkaline phosphatase and determination of its specific activity

Experiment 3 Effects of substrates concentration of enzymatic activity： determination of Km of alkaline phosphatase

Experiment 4 Inhibition effect of phosphate on alkaline phosphatase

Experiment 5 Analysis of lactic dehydrogenase isoenzyme

Chapter 9 Substance metabolism

Experiment 1 Extraction and identification of liver glycogen

Experiment 2 Determination of alanine transaminase activity （Reitman-Frankels'method）

Experiment 3 Enzyme catalyzed transamination of amino acids detected by paper chromatography

Chapter 10 Basic experiment of molecular biology

Experiment 1 Preparation of the genomic DNA in the eukaryotic cells

ExPeriment 2 Determination of the concentration and purity of DNA by spectroscopy

Experiment 3 Agarose gel electrophoresis of nucleic acids

Experiment 4 Polymerase chain reaction （PCR）

Experiment 5 Isolation and purification of the DNA fragments

Experiment 6 DNA ligation——constructing recombinant plasmids

Experiment 7 Preparation of E.coli competent cells

Experiment 8 Transformation of Ca2+ competent ceils

Experiment 9 The screening of recombinant DNA

Experiment 10 Minipreparation of plasmid DNA

Experiment 11 Digestion of plasmid DNA with restriction endonucleases

Experiment 12 Southern blot

Experiment 13 Expression of exogenous gene in prokaryotic cells

References

Appendix

Appendix I The calculation table of ammonium sulfate saturation

Appendix II Common buffer solution preparation

Appendix III The relative molecular mass and pI of protein

Appendix IV Common data of chromatography

Appendix V The conversion between centrifugation speed of centrifugation force

《Biochemistry and Molecular Biology Laboratory Course生物化学与分子生物学实验教程（英文版）》针对生物化学教学及研究中涉及的三类主要的生物分子一一蛋白质、酶和核酸，根据生物大分子的理伦性质，结合实用的电泳技术、层次技术、离心技术、分子克隆技术等，综合系统地介绍生物大分子的分离、纯化、含量测定、分析鉴定和应用，并结合教学中物质代谢、酶促反应动力学的系统分析，设计实验并融入相关实验技术和研究新进展，使学生在实验教学中掌握知识、技能的同时，体会对科研方法的选择与评价。
　　《Biochemistry and Molecular Biology Laboratory Course生物化学与分子生物学实验教程（英文版）》内容新颖、科学、实用性强，可作为高等医学院校本科学生、留学生及研究生教材使用，也可供有关的教学及科研人员参考。