24小时热门版块排行榜

返回列表

【奖励】本帖被评价103次，作者zqa224增加金币 82.2 个

zqa224

新虫 (初入文坛)

应助: 1 (幼儿园)
金币: 30.2
帖子: 17
在线: 10.6小时
虫号: 271991

[资源] 更新到Chapter 29的Current Protocols in Molecular Biology（分子克隆冷泉港出品）

目录：

Chapter 1 Escherichia coli, Plasmids, and BacteriophagesIntroduction
Section I Escherichia coliUnit 1.1 Media Preparation and Bacteriological Tools
Unit 1.2 Growth in Liquid Media
Unit 1.3 Growth on Solid Media
Unit 1.4 Selected Topics from Classical Bacterial Genetics
Section II Vectors Derived from PlasmidsUnit 1.5 Introduction to Plasmid Biology
Unit 1.6 Minipreps of Plasmid DNA
Unit 1.7 Large-Scale Preparation of Plasmid DNA
Unit 1.8 Introduction of Plasmid DNA into Cells
Section III Vectors Derived from Lambda and Related BacteriophagesUnit 1.9 Introduction to Lambda Phages
Unit 1.10 Lambda as a Cloning Vector
Unit 1.11 Plating Lambda Phage to Generate Plaques
Unit 1.12 Growing Lambda-Derived Vectors
Unit 1.13 Preparing Lambda DNA from Phage Lysates
Section IV Vectors Derived from Filamentous PhagesUnit 1.14 Introduction to Vectors Derived from Filamentous
Phages
Unit 1.15 Preparing and Using M13-Derived Vectors
Section V Specialized TechniquesUnit 1.16 Recombineering: Genetic Engineering in Bacteria Using Homologous
Recombination
Unit 1.17 E. coli Genome Manipulation by P1 Transduction
Chapter 2 Preparation and Analysis of DNAIntroduction
Section I Manipulation of DNAUnit 2.1A Purification and Concentration of DNA from Aqueous Solutions
Unit 2.1B Purification of DNA by Anion-Exchange Chromatography
Unit 2.2 Preparation of Genomic DNA from Mammalian Tissue
Unit 2.3 Preparation of Genomic DNA from Plant Tissue
Unit 2.4 Preparation of Genomic DNA from Bacteria
Section II Resolution and Recovery of Large DNA FragmentsUnit 2.5A Agarose Gel Electrophoresis
Unit 2.5B Pulsed-Field Gel Electrophoresis
Unit 2.6 Isolation and Purification of Large DNA Restriction Fragments from Agarose Gels
Section III Resolution and Recovery of Small DNA FragmentsUnit 2.7 Separation of Small DNA Fragments by
Conventional Gel Electrophoresis
Unit 2.8 Capillary Electrophoresis of DNA
Section IV Analysis of DNA Sequences by Blotting and HybridizationUnit 2.9A Southern Blotting
Unit 2.9B Dot and Slot Blotting of DNA
AUnit 2.10 Hybridization Analysis of DNA Blots
Section V Synthesis and Purification of OligonucleotidesUnit 2.11 Synthesis and Purification of Oligonucleotides
Unit 2.12 Purification of Oligonucleotides Using Denaturing Polyacrylamide Gel Electrophoresis
Chapter 3 Enzymatic Manipulation of DNA and RNAIntroduction
Section I Restriction EndonucleasesUnit 3.1 Digestion of DNA with Restriction Endonucleases
Section II Restriction MappingUnit 3.2 Mapping by Multiple Endonuclease Digestions
Unit 3.3 Mapping by Partial Endonuclease Digestions
Section III Enzymes for Modifying and Radioactively Labeling Nucleic AcidsUnit 3.4 Reagents and
Radioisotopes Used to Manipulate Nucleic Acids
Unit 3.5 DNA-Dependent DNA Polymerases
Unit 3.6 Template-Independent DNA Polymerases
Unit 3.7 RNA-Dependent DNA Polymerases
AUnit 3.8 DNA-Dependent RNA Polymerases
Unit 3.9 DNA-Independent RNA Polymerases
Unit 3.10 Phosphatases and Kinases
Unit 3.11 Exonucleases
Unit 3.12 Endonucleases
Unit 3.13 Ribonucleases
Unit 3.14 DNA Ligases
Unit 3.15 RNA Ligases
Section IV Construction of Hybrid DNA MoleculesUnit 3.16 Subcloning of DNA Fragments
Unit 3.17 Constructing Recombinant DNA Molecules by PCR
Section V Specialized ApplicationsUnit 3.18 Labeling and Colorimetric Detection of Nonisotopic Probes
Unit 3.19 Chemiluminescent Detection of Nonisotopic Probes
Unit 3.20 Recombinational Cloning
Chapter 4 Preparation and Analysis of RNAIntroduction
Section I Preparation of RNA from Eukaryotic and Prokaryotic CellsUnit 4.1 Preparation of Cytoplasmic RNA
from Tissue Culture Cells
Unit 4.2 Guanidine Methods for Total RNA Preparation
AUnit 4.3 Phenol/SDS Method for Plant RNA Preparation
Unit 4.4 Preparation of Bacterial RNA
Unit 4.5 Preparation of Poly(A)+ RNA
Section II Analysis of RNA Structure and SynthesisUnit 4.6 S1 Analysis of Messenger RNA Using
Single-Stranded DNA Probes
Unit 4.7 Ribonuclease Protection Assay
Unit 4.8 Primer Extension
Unit 4.9 Analysis of RNA by Northern and Slot Blot Hybridization
Unit 4.10 Identification of Newly Transcribed RNA
Chapter 5 Construction of Recombinant DNA LibrariesIntroduction
Section I Overview of Recombinant DNA LibrariesUnit 5.1 Genomic DNA Libraries
Unit 5.2 cDNA Libraries
Section II Preparation of Insert DNA from Genomic DNAUnit 5.3 Size Fractionation Using Sucrose Gradients
Unit 5.4 Size Fractionation Using Agarose Gels
Section III Preparation of Insert DNA from Messenger RNAUnit 5.5 Conversion of mRNA into Double-Stranded
cDNA
Unit 5.6 Ligation of Linkers or Adapters to Double-Stranded cDNA
Section IV Production of Genomic DNA and cDNA LibrariesUnit 5.7 Production of a Genomic DNA Library
Unit 5.8A Production of a Complete cDNA Library
Unit 5.9 Construction of Bacterial Artificial Chromosome (BAC/PAC) Libraries
Section V Amplification of Transformed or Packaged LibrariesUnit 5.10 Amplification of a Bacteriophage Library
Unit 5.11 Amplification of Cosmid and Plasmid Libraries
Chapter 6 Screening of Recombinant DNA LibrariesIntroduction
Section I Plating Libraries and Transfer to Filter MembranesUnit 6.1 Plating and Transferring Bacteriophage
Libraries
Unit 6.2 Plating and Transferring Cosmid and Plasmid Libraries
Section II Hybridization with Radioactive ProbesUnit 6.3 Using DNA Fragments as Probes
Unit 6.4 Using Synthetic Oligonucleotides as Probes
Section III Purification of Bacteriophage, Cosmid, and Plasmid ClonesUnit 6.5 Purification of Bacteriophage
Clones
Unit 6.6 Purification of Cosmid and Plasmid Clones
Section IV Screening with AntibodiesUnit 6.7 Immunoscreening of Fusion Proteins Produced in Lambda Plaques
Unit 6.8 Immunoscreening after Hybrid Selection and Translation
Section V Yeast Artificial Chromosome LibrariesUnit 6.9 Overview of Strategies for Screening YAC Libraries and
Analyzing YAC Clones
Unit 6.10 Analysis of Isolated YAC Clones
Section VI Specialized Strategies for Screening LibrariesUnit 6.11 Use of Monoclonal Antibodies for Expression
Cloning
Unit 6.12 Recombination-Based Assay (RBA) for Screening Bacteriophage Lambda Libraries
Chapter 7 DNA SequencingIntroduction
Unit 7.1 Overview of DNA Sequencing Strategies
Unit 7.2 Constructing Nested Deletions for Use in DNA Sequencing
Unit 7.3 Preparation of Templates for DNA Sequencing
Unit 7.4A DNA Sequencing by the Dideoxy Method
Unit 7.4B Dideoxy DNA Sequencing with Chemiluminescent Detection
Unit 7.5 DNA Sequencing by the Chemical Method
Unit 7.6 Denaturing Gel Electrophoresis for Sequencing
Unit 7.7 Computer Manipulation of DNA and Protein Sequences
Unit 7.8 Polony DNA Sequencing
Chapter 8 Mutagenesis of Cloned DNAIntroduction
Unit 8.1 Oligonucleotide-Directed Mutagenesis without Phenotypic Selection
Unit 8.2A Mutagenesis with Degenerate Oligonucleotides: Creating Numerous Mutations in a Small DNA
Sequence
Unit 8.2B Gene Synthesis: Assembly of Target Sequences Using Mutually Priming Long Oligonucleotides
Unit 8.3 Random Mutagenesis by PCR
Unit 8.4 Linker-Scanning Mutagenesis of DNA
Unit 8.5 Directed Mutagenesis Using the Polymerase Chain Reaction
Chapter 9 Introduction of DNA into Mammalian CellsIntroduction
Section I Transfection of DNA into Eukaryotic CellsUnit 9.1 Calcium Phosphate Transfection
Unit 9.2 Transfection Using DEAE-Dextran
Unit 9.3 Transfection by Electroporation
Unit 9.4 Transfection of Cultured Eukaryotic Cells Using Cationic Lipid Reagents
Unit 9.5 Selection of Transfected Mammalian Cells
Section II Uses of Fusion Genes in Mammalian TransfectionUnit 9.6 Overview of Genetic Reporter Systems
Unit 9.7A Isotopic Assays for Reporter Gene Activity
Unit 9.7B Nonisotopic Assays for Reporter Gene Activity
Unit 9.7C Use of the A. Victoria Green Fluorescent Protein to Study Protein Dynamics in Vivo
Unit 9.8 Direct Analysis of RNA after Transfection
Section III Transduction of Genes Using Retrovirus VectorsUnit 9.9 Overview of the Retrovirus Transduction
System
Unit 9.10 Preparation of a Specific Retrovirus Producer Cell Line
Unit 9.11 Transient Transfection Methods for Preparation of High-Titer Retroviral Supernatants
Unit 9.12 Large-Scale Preparation and Concentration of Retrovirus Stocks
Unit 9.13 Detection of Helper Virus in Retrovirus Stocks
Unit 9.14 Retrovirus Infection of Cells In Vitro and In Vivo
Section IV Inactivation of Genes in Mammalian CellsUnit 9.15 Human Somatic Cell Gene Targeting
Chapter 10 Analysis of ProteinsIntroduction
Section I Quantitation of ProteinsUnit 10.1A Spectrophotometric and Colorimetric Determination of Protein
Concentration
Unit 10.1B Quantitative Amino Acid Analysis
Section II Electrophoretic Separation of ProteinsUnit 10.2A One-Dimensional SDS Gel Electrophoresis of
Proteins
Unit 10.2B One-Dimensional Electrophoresis Using Nondenaturing Conditions
Unit 10.3 Two-Dimensional Gel Electrophoresis Using the ISO-DALT System
Unit 10.4 Two-Dimensional Gel Electrophoresis
Unit 10.5 Overview of Digital Electrophoresis Analysis
Section III Detection of ProteinsUnit 10.6 Staining Proteins in Gels
Unit 10.7 Detection of Proteins on Blot Transfer Membranes
Unit 10.8 Immunoblotting and Immunodetection
Section IV Purification of Proteins by Conventional ChromatographyUnit 10.9 Gel-Filtration Chromatography
Unit 10.10 Ion-Exchange Chromatography
Unit 10.11A Immunoaffinity Chromatography
Unit 10.11B Metal-Chelate Affinity Chromatography
Section V Purification of Proteins by High‑Performance Liquid ChromatographyUnit 10.12 HPLC of Peptides
and Proteins: Preparation and System Set-Up
Unit 10.13 HPLC of Peptides and Proteins: Standard Operating Conditions
Unit 10.14 Reversed-Phase Isolation of Peptides
Section VI Specialized ApplicationsUnit 10.15 Purification of Recombinant Proteins and Study of Protein
Interaction by Epitope Tagging
Unit 10.16 Immunoprecipitation
Unit 10.17 Synthesizing Proteins In Vitro by Transcription and Translation of Cloned Genes
Unit 10.18 Metabolic Labeling with Amino Acids
Unit 10.19 Isolation of Proteins for Microsequence Analysis
Unit 10.20 Capillary Electrophoresis of Proteins and Peptides
Unit 10.21 Overview of Peptide and Protein Analysis by Mass Spectrometry
Unit 10.22 Protein Identification and Characterization by Mass Spectrometry
Unit 10.23 Difference Gel Electrophoresis (DIGE) Using CyDye DIGE Fluor Minimal Dyes
Unit 10.24 Solution Radioimmunoassay of Proteins and Peptides
Chapter 11 ImmunologyIntroduction
Section I ImmunoassaysUnit 11.1 Conjugation of Enzymes to Antibodies
Unit 11.2 Enzyme-Linked Immunosorbent Assays (ELISA)
Unit 11.3 Isotype Determination of Antibodies
Section II Preparation of Monoclonal AntibodiesUnit 11.4 Immunization of Mice
Unit 11.5 Preparation of Myeloma Cells
Unit 11.6 Preparation of Mouse Feeder Cells for Fusion and Cloning
Unit 11.7 Fusion of Myeloma Cells with Immune Spleen Cells
Unit 11.8 Cloning of Hybridoma Cell Lines by Limiting Dilution
Unit 11.9 Freezing and Recovery of Hybridoma Cell Lines
Unit 11.10 Production of Monoclonal Antibody Supernatant and Ascites Fluid
Unit 11.11 Purification of Monoclonal Antibodies
Section III Preparation of Polyclonal AntiseraUnit 11.12 Production of Polyclonal Antisera
Unit 11.13 In Vitro Antibody Production
Unit 11.14 Purification of Immunoglobulin G Fraction from Antiserum, Ascites Fluid, or Hybridoma Supernatant
Section IV Preparation of Antipeptide AntibodiesUnit 11.15 Introduction to Peptide Synthesis
Unit 11.16 Synthetic Peptides for Production of Antibodies that Recognize Intact Proteins
Section V Determination of Specific Antibody Titer and IsotypeUnit 11.17 Determination of the Specific Antibody
Titer
Section VI Preparation and Use of Specialized AntibodiesUnit 11.18 Identification of Polyol-Responsive
Monoclonal Antibodies for Use in Immunoaffinity Chromatography
Chapter 12 DNA‑Protein InteractionsIntroduction
Unit 12.1 Preparation of Nuclear and Cytoplasmic Extracts from Mammalian Cells
Unit 12.2 Mobility Shift DNA-Binding Assay Using Gel Electrophoresis
Unit 12.3 Methylation and Uracil Interference Assays for Analysis of Protein-DNA Interactions
Unit 12.4 DNase I Footprint Analysis of Protein-DNA Binding
Unit 12.5 UV Crosslinking of Proteins to Nucleic Acids
Unit 12.6 Purification of DNA-Binding Proteins Using Biotin/Streptavidin Affinity Systems
Unit 12.7 Detection, Purification, and Characterization of cDNA Clones Encoding DNA-Binding Proteins
Unit 12.8 Rapid Separation of Protein-Bound DNA from Free DNA Using Nitrocellulose Filters
Unit 12.9 Analysis of DNA-Protein Interactions Using Proteins Synthesized In Vitro from Cloned Genes
Unit 12.10 Purification of Sequence-Specific DNA-Binding Proteins by Affinity Chromatography
Unit 12.11 Determination of Protein-DNA Sequence Specificity by PCR-Assisted Binding-Site Selection
Unit 12.12 Yeast One-Hybrid Screening for DNA-Protein Interactions
Chapter 13 YeastIntroduction
Section I Basic Techniques of Yeast GeneticsUnit 13.1 Preparation of Yeast Media
Unit 13.2 Growth and Manipulation of Yeast
Unit 13.3 Genome-Wide Transposon Mutagenesis in Yeast
Unit 13.3B EMS and UV Mutagenesis in Yeast
Section II Yeast VectorsUnit 13.4 Yeast Cloning Vectors and Genes
Unit 13.6 Yeast Vectors and Assays for Expression of Cloned Genes
Section III Manipulation of Yeast GenesUnit 13.7 Introduction of DNA into Yeast Cells
Unit 13.8 Cloning Yeast Genes by Complementation
Unit 13.9 Manipulation of Plasmids from Yeast Cells
Unit 13.10 Manipulation of Cloned Yeast DNA
Section IV Preparation of Yeast DNA, RNA, and ProteinsUnit 13.11 Preparation of Yeast DNA
Unit 13.12 Preparation of Yeast RNA
Unit 13.13 Preparation of Protein Extracts from Yeast
Section V Schizosaccharomyces pombeUnit 13.14 Overview of Schizosaccharomyces pombe
Unit 13.15 S. pombe Strain Maintenance and Media
Unit 13.16 Growth and Manipulation of S. pombe
Unit 13.17 Introduction of DNA into S. pombe Cells
Chapter 14 In Situ Hybridization and ImmunohistochemistryIntroduction
Unit 14.1 Fixation, Embedding, and Sectioning of Tissues, Embryos, and Single Cells
Unit 14.2 Cryosectioning
Unit 14.3 In Situ Hybridization to Cellular RNA
Unit 14.4 Detection of Hybridized Probe
Unit 14.5 Counterstaining and Mounting of Autoradiographed In Situ Hybridization Slides
Unit 14.6 Immunohistochemistry
Unit 14.7 In Situ Hybridization and Detection Using Nonisotopic Probes
Unit 14.8 In Situ Polymerase Chain Reaction and Hybridization to Detect Low-Abundance Nucleic Acid Targets
Unit 14.9 Whole-Mount In Situ Hybridization and Detection of RNAs in Vertebrate Embryos and Isolated Organs
Unit 14.10 Principles and Application of Fluorescence Microscopy
Unit 14.11 Basic Confocal Microscopy
Unit 14.12 Measurement of In Situ Hybridization
Unit 14.13 Morphological, Biochemical, and Flow Cytometric Assays of Apoptosis
Unit 14.14 Whole-Mount Histochemical Detection of -Galactosidase Activity
Unit 14.15 Overview of Image Analysis, Image Importing, and Image Processing using Freeware
Unit 14.16 Three-Dimensional Reconstruction of Tissues
Unit 14.17 Using CellProfiler for Automatic Identification and Measurement of Biological Objects in Images
Chapter 15 The Polymerase Chain ReactionIntroduction
Unit 15.1 Enzymatic Amplification of DNA by PCR: Standard Procedures and Optimization
Unit 15.2 Direct DNA Sequencing of PCR Products
Unit 15.3 Ligation-Mediated PCR for Genomic Sequencing and Footprinting
Unit 15.4 Molecular Cloning of PCR Products
Unit 15.5 Enzymatic Amplification of RNA by PCR (RT-PCR)
Unit 15.6 cDNA Amplification Using One-Sided (Anchored) PCR
Unit 15.7 Quantitation of Rare DNAs by PCR
Unit 15.8 High-Throughput Real-Time Quantitative Reverse Transcription PCR
Chapter 16 Protein ExpressionIntroduction
Section I Expression of Proteins in Escherichia coliUnit 16.1 Overview of Protein Expression in E. coli
Unit 16.2 Expression Using the T7 RNA Polymerase/Promoter System
Unit 16.3 Expression Using Vectors with Phage Regulatory Sequences
Unit 16.4A Introduction to Expression by Fusion Protein Vectors
Unit 16.4B Enzymatic and Chemical Cleavage of Fusion Proteins
Unit 16.5 Expression and Purification of lacZ and trpE Fusion Proteins
Unit 16.6 Expression and Purification of Maltose-Binding Protein Fusions
Unit 16.7 Expression and Purification of Glutathione-S-Transferase Fusion Proteins
Unit 16.8 Expression and Purification of Thioredoxin Fusion Proteins
Section II Expression of Proteins in Insect Cells Using Baculovirus VectorsUnit 16.9 Overview of the Baculovirus
Expression System
Unit 16.10 Maintenance of Insect Cell Cultures and Generation of Recombinant Baculoviruses
Unit 16.11 Expression and Purification of Recombinant Proteins Using the Baculovirus System
Section III Expression of Proteins in Mammalian CellsUnit 16.12 Transient Expression of Proteins Using COS
Cells
Unit 16.13 Expression and Purification of Epitope-Tagged Multisubunit Protein Complexes from Mammalian
Cells
Unit 16.14 Inducible Gene Expression Using an Autoregulatory, Tetracycline-Controlled System
Unit 16.15 Overview of the Vaccinia Virus Expression System
Unit 16.16 Preparation of Cell Cultures and Vaccinia Virus Stocks
Unit 16.17 Generation of Recombinant Vaccinia Viruses
Unit 16.18 Characterization of Recombinant Vaccinia Viruses and Their Products
Unit 16.19 Gene Expression Using the Vaccinia Virus/T7 RNA Polymerase Hybrid System
Unit 16.20 Expression of Proteins Using Semliki Forest Virus Vectors
Unit 16.21 Overview of the HIV-1 Lentiviral Vector System
Unit 16.22 Generation of HIV-1-Based Lentiviral Vector Particles
Unit 16.23 Amplification Using CHO Cell Expression Vectors
Unit 16.24 Helper-Dependent Adenoviral Vectors
Unit 16.25 Production of Recombinant Adeno-Associated Viral Vectors for In Vitro and In Vivo Use
Chapter 17 Preparation and Analysis of GlycoconjugatesIntroduction
Section I Special Considerations for Glycoproteins and Their PurificationUnit 17.1 Special Considerations for
Glycoproteins and Their Purification
Unit 17.2 Special Considerations for Proteoglycans and Glycosaminoglycans and Their Purification
Unit 17.3 Special Considerations for Glycolipids and Their Purification
Section II Detection of Saccharides on GlycoconjugatesUnit 17.4 Metabolic Radiolabeling of Animal Cell
Glycoconjugates
Unit 17.5 Chemical Labeling of Carbohydrates by Oxidation and Sodium Borohydride Reduction
Unit 17.6 Detection and Analysis of Proteins Modified by O-Linked N-Acetylglucosamine
Unit 17.7 Lectin Analysis of Proteins Blotted onto Filters
Unit 17.8 Detection of Glycophospholipid Anchors on Proteins
Unit 17.9 Direct Chemical Analysis of Glycoconjugates for Carbohydrates
Unit 17.10A Inhibition of N-Linked Glycosylation
Unit 17.10B Inhibition of Glycolipid Biosynthesis
Unit 17.11 Synthetic Glycosides as Primers of Oligosaccharide Biosynthesis and Inhibitors of Glycoprotein and
Proteoglycan Assembly
Section III Release of Saccharides from GlycoconjugatesUnit 17.12 Sialidases
Unit 17.13A Endoglycosidase and Glycoamidase Release of N-Linked Oligosaccharides
Unit 17.13B Analysis of Glycosaminoglycans with Polysaccharide Lyases
Unit 17.14A Preparation of Glycopeptides
Unit 17.14B Detection of Individual Glycosylation Sites on Glycoproteins
Unit 17.15A -Elimination for Release of O-Linked Glycosaminoglycans from Proteoglycans
Unit 17.15B -Elimination for Release of O-GalNAc-Linked Oligosaccharides from Glycoproteins and
Glycopeptides
Unit 17.16 Acid Hydrolysis for Release of Monosaccharides
Unit 17.17A Enzymatic Release of Oligosaccharides from Glycolipids
Unit 17.17B Endo--Galactosidases and Keratanase
Section IV Analysis of Saccharides Released from GlycoconjugatesUnit 17.18 Analysis of Monosaccharides
Unit 17.19A Total Compositional Analysis by High-Performance Liquid Chromatography or Gas-Liquid
Chromatography
Unit 17.19B Composition of Labeled Monosaccharides from Glycosaminoglycans
Unit 17.20 Analysis of Oligosaccharide Negative Charge by Anion-Exchange Chromatography
Unit 17.21A HPLC Methods for the Fractionation and Analysis of Negatively Charged Oligosaccharides and
Gangliosides
Unit 17.21B Fractionation and Analysis of Neutral Oligosaccharides by HPLC
Unit 17.22A Nitrous Acid Degradation of Glycosaminoglycans
Unit 17.22B Analysis of Disaccharides and Tetrasaccharides Released from Glycosaminoglycans
Unit 17.23 Analysis of Sulfate Esters by Solvolysis or Hydrolysis
Chapter 18 Analysis of Protein PhosphorylationIntroduction
Unit 18.1 Overview of Protein Phosphorylation
Unit 18.2 Labeling Cultured Cells with 32Pi and Preparing Cell Lysates for Immunoprecipitation
Unit 18.3 Phosphoamino Acid Analysis
Unit 18.4 Analysis of Phosphorylation of Unlabeled Proteins
Unit 18.5 Detection of Phosphorylation by Enzymatic Techniques
Unit 18.6 Production of Antibodies That Recognize Specific Tyrosine-Phosphorylated Peptides
Unit 18.7 Assays of Protein Kinases Using Exogenous Substrates
Unit 18.8 Permeabilization Strategies to Study Protein Phosphorylation
Unit 18.9 Phosphopeptide Mapping and Identification of Phosphorylation Sites
Unit 18.10 Use of Protein Phosphatase Inhibitors
Unit 18.11 Design and Use of Analog-Sensitive Protein Kinases
Unit 18.12 The Detection of MAPK Signaling
Unit 18.13 Isolation of Phosphopeptides by Immobilized Metal Ion Affinity Chromatography
Chapter 19 Informatics for Molecular BiologistsIntroduction
Unit 19.1 Internet Basics for Biologists
Unit 19.2 Sequence Databases: Integrated Information Retrieval and Data Submission
Unit 19.3 Sequence Similarity Searching Using the BLAST Family of Programs
Unit 19.4 Protein Databases on the Internet
Unit 19.5 Basic Protein Sequence Analysis
Unit 19.6 Analysis and Management of Microarray Gene Expression Data
Chapter 20 Analysis of Protein InteractionsIntroduction
Unit 20.1 Interaction Trap/Two-Hybrid System to Identify Interacting Proteins
Unit 20.2 Affinity Purification of Proteins Binding to GST Fusion Proteins
Unit 20.3 Phage-Based Expression Cloning to Identify Interacting Proteins
Unit 20.4 Surface Plasmon Resonance for Measurements of Biological Interest
Unit 20.5 Detection of Protein-Protein Interactions by Coprecipitation
Unit 20.6 Identification of Protein Interactions by Far Western Analysis
Unit 20.7 Two-Hybrid Dual Bait System
Unit 20.8 Interaction Trap/Two-Hybrid System to Identify Loss-of-Interaction Mutant Proteins
Chapter 21 Chromatin Assembly and AnalysisIntroduction
Unit 21.1 Micrococcal Nuclease Analysis of Chromatin Structure
Unit 21.2 Separation of Histone Variants and Post-Translationally Modified Isoforms by Triton/Acetic Acid/Urea
Polyacrylamide Gel Electrophoresis
Unit 21.3 Chromatin Immunoprecipitation for Determining the Association of Proteins with Specific Genomic
Sequences In Vivo
Unit 21.4 DNase I and Hydroxyl Radical Characterization of Chromatin Complexes
Unit 21.5 Isolation of Histones and Nucleosome Cores from Mammalian Cells
Unit 21.6 Assembly of Nucleosomal Templates by Salt Dialysis
Unit 21.7 Chromatin Assembly Using Drosophila Systems
Unit 21.8 Analysis of Protein Co-Occupancy by Quantitative Sequential Chromatin Immunoprecipitation
Unit 21.9 Defining In Vivo Targets of Nuclear Proteins by Chromatin Immunoprecipitation and Microarray
Analysis
Unit 21.10 Identifying Chromosomal Targets of DNA-Binding Proteins by Sequence Tag Analysis of Genomic
Enrichment (STAGE)
Unit 21.11 Mapping Chromatin Interactions by Chromosome Conformation Capture
Unit 21.12 Paired-End diTagging for Transcriptome and Genome Analysis
Unit 21.13 ChIP-chip for Genome-Wide Analysis of Protein Binding in Mammalian Cells
Unit 21.14 Chromosome Conformation Capture Carbon Copy Technology
Chapter 22 Nucleic Acid ArraysIntroduction
Unit 22.1 Overview of Nucleic Acid Arrays
Unit 22.2 Preparation of mRNA for Expression Monitoring
Unit 22.3 Profiling Human Gene Expression with cDNA Microarrays
Unit 22.4 Overview of mRNA Expression Profiling Using Microarrays
Unit 22.5 Pattern Discovery in Expression Profiling Data
Chapter 23 Manipulating the Mouse GenomeIntroduction
Unit 23.1 Overview of Gene Targeting by Homologous Recombination
Unit 23.2 Mouse Embryo Fibroblast (MEF) Feeder Cell Preparation
Unit 23.3 Mouse Embryonic Stem (ES) Cell Culture
Unit 23.4 Mouse Embryonic Stem (ES) Cell Isolation
Unit 23.5 Production of a Heterozygous Mutant Cell Line by Homologous Recombination (Single Knockout)
Unit 23.6 Production of a Homozygous Mutant Embryonic Stem Cell Line (Double Knockout)
Unit 23.7 Chimeric Mouse Production by Microinjection
Unit 23.8 Mouse Colony Management
Unit 23.9 Transgenic Mouse Production By Zygote Injection
Unit 23.10 Transgenic Mouse Colony Management
Unit 23.11 Modification and Production of BAC Transgenes
Unit 23.12 Regulation of Transgene Expression Using Tetracycline
Chapter 24 Generation and Use of Combinatorial LibrariesUnit 24.1 Overview of Receptors from Combinatorial
Nucleic Acid and Protein Libraries
Unit 24.2 Design, Synthesis, and Amplification of DNA Pools for Construction of Combinatorial Pools and
Libraries
Unit 24.3 In Vitro Selection of RNA Aptamers to a Protein Target by Filter Immobilization
Unit 24.4 Peptide Aptamers: Dominant “Genetic” Agents for Forward and Reverse Analysis of Cellular Processes
Unit 24.5 Protein Selection Using mRNA Display
Chapter 25 Discovery and Analysis of Differentially Expressed Genes in Single Cells and Cell
PopulationsIntroduction
Section A Nucleic Acid Amplification from Individual CellsUnit 25A.1 Laser Capture Microdissection
Unit 25A.2 Preparation of Single Cells from Solid Tissues for Analysis by PCR
Section B Molecular Methods for Discovery of Differentially Expressed GenesUnit 25B.1 Production of a
Subtracted cDNA Library
Unit 25B.2 PCR-Based Subtractive cDNA Cloning
Unit 25B.3 Differential Display of mRNA by PCR
Unit 25B.4 Restriction-Mediated Differential Display (RMDD)
Unit 25B.5 AFLP-Based Transcript Profiling
Unit 25B.6 Serial Analysis of Gene Expression (SAGE): Experimental Method and Data Analysis
Unit 25B.7 Representational Difference Analysis
Unit 25B.8 Gene Expression Analysis of a Single or Few Cells
Chapter 26 Gene SilencingIntroduction
Unit 26.1 Overview of RNA Interference and Related Processes
Unit 26.2 Gene Silencing by RNAi in Mammalian Cells
Unit 26.3 RNA Interference in Caenorhabditis Elegans
Unit 26.4 Cloning of Small RNA Molecules
Unit 26.5 RNA Interference in Cultured Drosophila Cells
Unit 26.6 RNAi in Transgenic Plants
Unit 26.7 Analysis of Small Endogenous RNAs
Unit 26.8 Using Morpholinos to Control Gene Expression
Chapter 27 RNA‑Protein InteractionsIntroduction
Unit 27.1 Agarose Gel Separation/Isolation of RNA-Protein Complexes
Unit 27.2 Identification of RNA Binding Proteins by UV Cross-Linking
Unit 27.3 Purification of Functional RNA-Protein Complexes using MS2-MBP
Unit 27.4 RNA Immunoprecipitation for Determining RNA-Protein Associations In Vivo
Chapter 28 Mammalian Cell CultureIntroduction
Unit 28.1 Preparation, Culture, and Immortalization of Mouse Embryonic Fibroblasts
Unit 28.2 Isolation and Immortalization of Lymphocytes
Unit 28.3 Establishment and Culture of Human Skin Fibroblasts
Chapter 29 Mouse PhenotypingIntroduction
Section A General Considerations in Mouse PhenotypingUnit 29A.1 Uses of Forward and Reverse Genetics in
Mice to Study Gene Function
Unit 29A.2 Minimizing Variation Due to Genotype and Environment
Unit 29A.3 Collection of Blood and Plasma from the Mouse
Unit 29A.4 Tissue Collection for Systematic Phenotyping in the Mouse
Section B Metabolic Exploration of the MouseUnit 29B.1 Evaluation of Energy Homeostasis
Unit 29B.2 Lipid and Bile Acid Analysis
Unit 29B.3 Evaluation of Glucose Homeostasis
Unit 29B.4 Histopathology in Mouse Metabolic Investigations
Appendix 1 Standard Measurements, Data, and Abbreviations1A Common Abbreviations
1B Useful Measurements and Data
1C Characteristics of Amino Acids
1D Characteristics of Nucleic Acids
1E Radioactivity
1F Safe Use of Radioisotopes
1G Centrifuges and Rotors
1H Safe Use of Hazardous Chemicals
1I Commonly Used Detergents
1J Common Conversion Factors
1K Compendium of Drugs Commonly Used in Molecular Biology Research
Appendix 2 Commonly Used Reagents and Equipment2 Commonly Used Reagents and Equipment
Appendix 3 Commonly Used Techniques in Biochemistry and Molecular Biology3A Detection and Quantitation
of Radiolabeled Proteins and DNA in Gels and Blots
3B Silanizing Glassware
3C Dialysis and Ultrafiltration
3D Quantitation of DNA and RNA with Absorption and Fluorescence Spectroscopy
3E Introduction of Restriction Enzyme Recognition Sequences by Silent Mutation
3F Techniques for Mammalian Cell Tissue Culture
3G Importing Biological Materials
3H Kinetic Assay Methods
3I Statistics for the Molecular Biologist: Group Comparisons
Appendix 4 Suppliers4 Selected Suppliers of Reagents and Equipment
Appendix 5 Vectors5 Vectors

回复此楼

» 本帖附件资源列表

欢迎监督和反馈：小木虫仅提供交流平台，不对该内容负责。
本内容由用户自主发布，如果其内容涉及到知识产权问题，其责任在于用户本人，如对版权有异议，请联系邮箱：xiaomuchong@tal.com
附件 1 : Current_Protocols_in_Molecular_Biology-all.pdf

2016-04-09 17:45:58, 46.15 M