生物化学与分子生物学实验教程

Part 1 Basic techniques of biochemistry and molecular biologyChapter 1 Basic techniques of preparation of biomacromoleculesSection 1 Salting outSection 2 Dialysis and ultrafiltrationChapter 2 SpectrophotometrySection 1 Basic principlesSection 2 The basic construction of spectrophotometersSection 3 Some home-made spectrophotometersChapter 3 ChromatographySection 1 Adsorption chromatographySection 2 Partition chromatographySection 3 Ion-exchange chromatographySection 4 Gel filtrationSection 5 Affinity chromatographyChapter 4 Electrophoretic techniquesSection 1 Basic principlesSection 2 Several types of commonly used electrophoresisChapter 5 Centrifugation techniquesSection 1 Basic principleSection 2 General description of centrifugationChapter 6 Genetic engineering and analytieal technology of gene expressionSection 1 Genetic engineeringSection 2 Polymerase chain reactionSection 3 Nucleic acid hybridizationSection 4 DNA sequencing technologyPart 2 Experiments of the biochemistry and molecular biologyChapter 7 Analysis of purification and physicochemical properties of proteinsExperiment 1 Colorimetric methods for protein determinationExperiment 2 Isolation and purification of serum y-globulinExperiment 3 Cellulose acetate membrane electrophoresis of serum proteinsExperiment 4 Determination of relative molecular mass of protein in SDS-PAGEChapter 8 Kinetics of enzyme-catalysed reactionsExperiment 1 The specificity of enzyme and effect of temperature， pH on the enzyme ActivityExperiment 2 Isolation of alkaline phosphatase and determination of its specific activityExperiment 3 Effects of substrates concentration of enzymatic activity： determination of Km of alkaline phosphataseExperiment 4 Inhibition effect of phosphate on alkaline phosphataseExperiment 5 Analysis of lactic dehydrogenase isoenzymeChapter 9 Substance metabolismExperiment 1 Extraction and identification of liver glycogenExperiment 2 Determination of alanine transaminase activity （Reitman-Frankels’method）Experiment 3 Enzyme catalyzed transamination of amino acids detected by paper chromatographyChapter 10 Basic experiment of molecular biologyExperiment 1 Preparation of the genomic DNA in the eukaryotic cellsExPeriment 2 Determination of the concentration and purity of DNA by spectroscopyExperiment 3 Agarose gel electrophoresis of nucleic acidsExperiment 4 Polymerase chain reaction （PCR）Experiment 5 Isolation and purification of the DNA fragmentsExperiment 6 DNA ligation——constructing recombinant plasmidsExperiment 7 Preparation of E.coli competent cellsExperiment 8 Transformation of Ca2+ competent ceilsExperiment 9 The screening of recombinant DNAExperiment 10 Minipreparation of plasmid DNAExperiment 11 Digestion of plasmid DNA with restriction endonucleasesExperiment 12 Southern blotExperiment 13 Expression of exogenous gene in prokaryotic cellsReferencesAppendixAppendix I The calculation table of ammonium sulfate saturationAppendix II Common buffer solution preparationAppendix III The relative molecular mass and pI of proteinAppendix IV Common data of chromatographyAppendix V The conversion between centrifugation speed of centrifugation force