Current Protocols in Molecular Biology

Current Protocols in Molecular Biology

Online ISBN: 9780471142720

DOI: 10.1002/0471142727

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Current Protocols in Molecular Biology Supplements

2014:    Supplement 106     Supplement 105
2013:    Supplement 104     Supplement 103     Supplement 102     Supplement 101
2012:    Supplement 100     Supplement 99     Supplement 98     Supplement 97
2011:    Supplement 96     Supplement 95     Supplement 94     Supplement 93
2010:    Supplement 92     Supplement 91     Supplement 90     Supplement 89
2009:    Supplement 88     Supplement 87     Supplement 86     Supplement 85
2008:    Supplement 84     Supplement 83     Supplement 82     Supplement 81
2007:    Supplement 80     Supplement 79     Supplement 78     Supplement 77    
2006:    Supplement 76     Supplement 75     Supplement 74     Supplement 73    
2005:    Supplement 72     Supplement 71     Supplement 70     Supplement 69        
2004:    Supplement 68     Supplement 67     Supplement 66     Supplement 65        
2003:    Supplement 64     Supplement 63     Supplement 62     Supplement 61        
2002:    Supplement 60     Supplement 59     Supplement 58     Supplement 57        
2001:    Supplement 56     Supplement 55     Supplement 54     Supplement 53        
2000:    Supplement 52     Supplement 51     Supplement 50     Supplement 49        
1999:    Supplement 48     Supplement 47     Supplement 46     Supplement 45        
1998:    Supplement 44     Supplement 43     Supplement 42     Supplement 41    
1997:    Supplement 40     Supplement 39     Supplement 38     Supplement 37    
1996:    Supplement 36     Supplement 35     Supplement 34     Supplement 33    
1995:    Supplement 32     Supplement 31     Supplement 30     Supplement 29    
1994:    Supplement 28     Supplement 27     Supplement 26     Supplement 25    
1993:    Supplement 24     Supplement 23     Supplement 22     Supplement 21    
1992:    Supplement 20     Supplement 19     Supplement 18     Supplement 17    
1991:    Supplement 16     Supplement 15     Supplement 14     Supplement 13    
1990:    Supplement 12     Supplement 11     Supplement 10     Supplement 9    
1989:    Supplement 8       Supplement 7       Supplement 6       Supplement 5    
1988:    Supplement 4       Supplement 3       Supplement 2       Supplement 1    
1987:    Core publication




Supplement 106, April 2014

Unit 1.16 Recombineering: Genetic Engineering in Bacteria Using Homologous Recombination
Abstract | Full Text: HTML PDF

Unit 4.21 Next-Generation Sequencing RNA-Seq Library Construction
Abstract | Full Text: HTML PDF

Unit 7.16 Genome-Wide Fitness and Genetic Interactions Determined by Tn-seq, a High-Throughput Massively Parallel Sequencing Method for Microorganisms
Abstract | Full Text: HTML PDF

Unit 28.4 Detection of Mycoplasma Contamination in Cell Cultures
Abstract | Full Text: HTML PDF

Unit 28.5 Eradication of Mycoplasma Contaminations from Cell Cultures
Abstract | Full Text: HTML PDF

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Supplement 105, January 2014

Unit 4.20 Purification of Radiolabeled RNA Products Using Denaturing Gel Electrophoresis
Abstract | Full Text: HTML PDF

Unit 14.23 Visualizing Genomes with Oligopaint FISH Probes
Abstract | Full Text: HTML PDF

Unit 15.12 Library Generation by Gene Shuffling
Abstract | Full Text: HTML PDF

Unit 15.13 Random-Primed, Phi29 DNA Polymerase–Based Whole Genome Amplification
Abstract | Full Text: HTML PDF

Unit 15.14 Loop-Mediated Isothermal Amplification for Detection of Nucleic Acids
Abstract | Full Text: HTML PDF

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Supplement 104, October 2013

Unit 7.13 Identification of Mutations in Zebrafish Using Next-Generation Sequencing
Abstract | Full Text: HTML PDF

Unit 7.14 Whole-Genome Amplification of Single-Cell Genomes for Next-Generation Sequencing
Abstract | Full Text: HTML PDF

Unit 7.15 Pyrosequencing: Powerful and Quantitative Sequencing Technology
Abstract | Full Text: HTML PDF

Unit 13.10C Construction of Mutant Alleles in Saccharomyces cerevisiae without Cloning: Overview and the Delitto Perfetto Method
Abstract | Full Text: HTML PDF

Unit 15.11 Helicase-Dependent Amplification of Nucleic Acids
Abstract | Full Text: HTML PDF

Unit 25B.11 RAMPAGE: Promoter Activity Profiling by Paired-End Sequencing of 5′-Complete cDNAs
Abstract | Full Text: HTML PDF

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Supplement 103, July 2013

Unit 4.16 RNA Isolation from Mammalian Samples
Abstract | Full Text: HTML PDF

Unit 4.17 Quantification of microRNA Expression with Next-Generation Sequencing
Abstract | Full Text: HTML PDF

Unit 4.18 Genome-Wide Annotation and Quantitation of Translation by Ribosome Profiling
Abstract | Full Text: HTML PDF

Unit 4.19 Ribosomal RNA Depletion for Efficient Use of RNA-Seq Capacity
Abstract | Full Text: HTML PDF

Unit 12.16 Engineering Customized TALE Nucleases (TALENs) and TALE Transcription Factors by Fast Ligation-Based Automatable Solid-Phase High-Throughput (FLASH) Assembly
Abstract | Full Text: HTML PDF

Unit 21.27 Genome-Scale Mapping of DNase I Hypersensitivity
Abstract | Full Text: HTML PDF

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Supplement 102, April 2013

Unit 2.14 DNA Isolation from Mammalian Samples
Abstract | Full Text: HTML PDF

Unit 4.15 Synthesis and Labeling of RNA In Vitro
Abstract | Full Text: HTML PDF

Unit 7.12 Exome Sequencing by Targeted Enrichment
Abstract | Full Text: HTML PDF

Unit 14.15 Basic Image Analysis and Manipulation in ImageJ
Abstract | Full Text: HTML PDF

Unit 21.26 A Detailed Protocol for Formaldehyde-Assisted Isolation of Regulatory Elements (FAIRE)
Abstract | Full Text: HTML PDF

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Supplement 101, January 2013

Unit 7.11 Multiplex Illumina Sequencing Using DNA Barcoding
Abstract | Full Text: HTML PDF

Unit 14.22 Using CellX to Quantify Intracellular Events
Abstract | Full Text: HTML PDF

Unit 18.20 Using Phospho-Motif Antibodies to Determine Kinase Substrates
Abstract | Full Text: HTML PDF

Unit 19.1 Detecting the Signatures of Adaptive Evolution in Protein-Coding Genes
Abstract | Full Text: HTML PDF

Unit 21.25 Capture Hybridization Analysis of RNA Targets (CHART)
Abstract | Full Text: HTML PDF

Unit 22.1 Overview of DNA Microarrays: Types, Applications, and Their Future
Abstract | Full Text: HTML PDF

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Supplement 100, October 2012

Unit 12.15 Engineering Designer Transcription Activator–Like Effector Nucleases (TALENs) by REAL or REAL-Fast Assembly
Abstract | Full Text: HTML PDF

Unit 14.18 Using Cell-ID 1.4 with R for Microscope-Based Cytometry
Abstract | Full Text: HTML PDF

Unit 21.24 ChIP-exo Method for Identifying Genomic Location of DNA-Binding Proteins with Near-Single-Nucleotide Accuracy
Abstract | Full Text: HTML PDF

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Supplement 99, July 2012

Unit 3.23 Design and Assembly of Large Synthetic DNA Constructs
Abstract | Full Text: HTML PDF

Unit 3.24 Enrichment of Error-Free Synthetic DNA Sequences by CEL I Nuclease
Abstract | Full Text: HTML PDF

Unit 3.25 Database of DNA Polymerases
Abstract | Full Text: HTML PDF

Unit 10.29 Biochemical Analysis of Protein SUMOylation
Abstract | Full Text: HTML PDF

Unit 10.30 Assays for Investigating deSUMOylation Enzymes
Abstract | Full Text: HTML PDF

Unit 21.23 Single Nucleotide Analysis of Cytosine Methylation by Whole-Genome Shotgun Bisulfite Sequencing
Abstract | Full Text: HTML PDF

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Supplement 98, April 2012

Unit 2.13 Preparation of Genomic DNA from Mammalian Sperm
Abstract | Full Text: HTML PDF

Unit 4.13 RASL-seq for Massively Parallel and Quantitative Analysis of Gene Expression
Abstract | Full Text: HTML PDF

Unit 4.14 Native Elongating Transcript Sequencing (NET-seq)
Abstract | Full Text: HTML PDF

Unit 14.21 Use of YouScope to Implement Systematic Microscopy Protocols
Abstract | Full Text: HTML PDF

Unit 18.19 Determining In Vivo Phosphorylation Sites Using Mass Spectrometry
Abstract | Full Text: HTML PDF

Unit 30.2 Targeted Metabolomics
Abstract | Full Text: HTML PDF

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Supplement 97, January 2012

Unit 10.2A One-Dimensional SDS Gel Electrophoresis of Proteins
Abstract | Full Text: HTML PDF

Unit 12.14 In Vitro Transcription and Immobilized Template Analysis of Preinitiation Complexes
Abstract | Full Text: HTML PDF

Unit 19.4 Protein Databases on the Internet
Abstract | Full Text: HTML PDF

Unit 23.15 Reprogramming of Mouse, Rat, Pig, and Human Fibroblasts into iPS Cells
Abstract | Full Text: HTML PDF

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Supplement 96, October 2011

Unit 3.11 Exonucleases
Abstract | Full Text: HTML PDF

Unit 7.0 Introduction and Historical Overview of DNA Sequencing
Abstract | Full Text: HTML PDF

Unit 7.1 Overview of DNA Sequencing Strategies
Abstract | Full Text: HTML PDF

Unit 7.2 “First Generation” Automated DNA Sequencing Technology
Abstract | Full Text: HTML PDF

Unit 7.5 High-Throughput Sequencing of PCR Products Tagged with Universal Primers Using 454 Life Sciences System
Abstract | Full Text: HTML PDF

Unit 12.13 Engineering Designer Nucleases with Customized Cleavage Specificities
Abstract | Full Text: HTML PDF

Unit 19.11 Phylogenetic Analysis of Protein Sequence Data Using the Randomized Axelerated Maximum Likelihood (RAXML) Program
Abstract | Full Text: HTML PDF

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Supplement 95, July 2011

Unit 15.10 Stem-Loop RT-qPCR for miRNAs
Abstract | Full Text: HTML PDF

Unit 17.6 Detection and Analysis of Proteins Modified by O-Linked N-Acetylglucosamine
Abstract | Full Text: HTML PDF

Unit 21.22 Simultaneous Single-Molecule Mapping of Protein-DNA Interactions and DNA Methylation by MAPit
Abstract | Full Text: HTML PDF

Unit 23.14 Recombineering-Based Procedure for Creating BAC Transgene Constructs for Animals and Cell Lines
Abstract | Full Text: HTML PDF

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Supplement 94, April 2011

Unit 3.14 DNA Ligases
Abstract | Full Text: HTML PDF

Unit 3.22 Gene and Genome Construction in Yeast
Abstract | Full Text: HTML PDF

Unit 4.12 Construction of Normalized RNA-seq Libraries for Next-Generation Sequencing Using the Crab Duplex-Specific Nuclease
Abstract | Full Text: HTML PDF

Unit 10.28 The Combined Use of the Thermofluor Assay and ThermoQ Analytical Software for the Determination of Protein Stability and Buffer Optimization as an Aid in Protein Crystallization
Abstract | Full Text: HTML PDF

Unit 21.21 DamIP: Using Mutant DNA Adenine Methyltransferase to Study DNA-Protein Interactions In Vivo
Abstract | Full Text: HTML PDF

Unit 25B.10 Digital Multiplexed Gene Expression Analysis Using the NanoString nCounter System
Abstract | Full Text: HTML PDF

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Supplement 93, January 2011

Unit 3.12 Endonucleases
Abstract | Full Text: HTML PDF

Unit 13.10B Conditional Depletion of Nuclear Proteins by the Anchor Away System
Abstract | Full Text: HTML PDF

Unit 25B.9 Quantification of Gene Transcripts with Deep Sequencing Analysis of Gene Expression (DSAGE) Using 1 to 2 μg Total RNA
Abstract | Full Text: HTML PDF

Appendix 3D Quantitation of DNA and RNA with Absorption and Fluorescence Spectroscopy
Abstract | Full Text: HTML PDF

Appendix 3J Microvolume Quantitation of Nucleic Acids
Abstract | Full Text: HTML PDF

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Supplement 92, October 2010

Unit 7.10 Single Molecule Sequencing with a HeliScope Genetic Analysis System
Abstract | Full Text: HTML PDF

Unit 9.3 Transfection of Electroporation
Abstract | Full Text: HTML PDF

Unit 13.2B Drug-Induced Permeabilization of S. cerevisiae
Abstract | Full Text: HTML PDF

Unit 14.20 Computer Control of Microscopes Using μManager
Abstract | Full Text: HTML PDF

Unit 18.18 Serine/Threonine Protein Phosphatase Assays
Abstract | Full Text: HTML PDF

Unit 21.20 Genome-Wide Location Analysis by Pull Down of In Vivo Biotinylated Transcription Factors
Abstract | Full Text: HTML PDF

Appendix 6 Pathway Modulators and Inhibitors
Abstract | Full Text: HTML PDF

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Supplement 91, July 2010

Unit 7.9 Bisulfite Sequencing of DNA
Abstract | Full Text: HTML PDF

Unit 18.14 Analysis of Serine-Threonine Kinase Specificity Using Arrayed Positional Scanning Peptide Libraries
Abstract | Full Text: HTML PDF

Unit 18.15 Visualization of Kinase Activity with FRET-Based Activity Biosensors
Abstract | Full Text: HTML PDF

Unit 18.16 Analysis of Protein Tyrosine Phosphatases and Substrates
Abstract | Full Text: HTML PDF

Unit 18.17 Fluorescent Peptide Assays for Protein Kinases
Abstract | Full Text: HTML PDF

Unit 21.19 ChIP-Seq: A Method for Global Identification of Regulatory Elements in the Genome
Abstract | Full Text: HTML PDF

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Supplement 90, April 2010

Unit 5.12 Normalizing cDNA Libraries
Abstract | Full Text: HTML PDF

Unit 10.26 Isolation of Proteins by Heat-Induced Extraction from Formalin-Fixed, Paraffin-Embedded Tissue and Preparation of Tryptic Peptides for Mass Spectrometric Analysis
Abstract | Full Text: HTML PDF

Unit 10.27 Extraction of Proteins from Formalin-Fixed, Paraffin-Embedded Tissue Using the Qproteome Extraction Technique and Preparation of Tryptic Peptides for Liquid Chromatography/Mass Spectrometry Analysis
Abstract | Full Text: HTML PDF

Unit 21.18 Installation of Site-Specific Methylation into Histones Using Methyl Lysine Analogs
Abstract | Full Text: HTML PDF

Unit 30.1 Untargeted Metabolomics
Abstract | Full Text: HTML PDF

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Supplement 89, January 2010

Unit 4.11 RNA-Seq: A Method for Comprehensive Transcriptome Analysis
Abstract | Full Text: HTML PDF

Unit 17.13A Endoglycosidase and Glycoamidase Release from N-Linked Glycans
Abstract | Full Text: HTML PDF

Unit 19.10 Galaxy: A Web-Based Genome Analysis Tool for Experimentalists
Abstract | Full Text: HTML PDF

Unit 21.15 Chromatin Interaction Analysis Using Paired-End Tag Sequence
Abstract | Full Text: HTML PDF

Unit 21.16 Mapping Networks of Protein-Mediated Physical Interactions Between Chromatin Elements
Abstract | Full Text: HTML PDF

Unit 21.17 Methylation-Sensitive Single-Molecule Analysis of Chromatin Structure
Abstract | Full Text: HTML PDF

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Supplement 88, October 2009

Unit 10.25 Preparation of Proteins and Peptides for Mass Spectrometry Analysis in a Bottom-Up Proteomics Workflow
Abstract | Full Text: HTML PDF

Unit 15.9 Heat-Activatable Primers for Hot-Start PCR and Hot-Start One-Step RT-PCR: Endpoint and Real-Time Experiments
Abstract | Full Text: HTML PDF

Unit 19.9 The UCSC Genome Browser: What Every Molecular Biologist Should Know
Abstract | Full Text: HTML PDF

Unit 24.2 Design, Synthesis, and Amplification of DNA Pools for In Vitro Selection
Abstract | Full Text: HTML PDF

Unit 24.3 In Vitro Selection of RNA Aptamers to a Protein Target by Filter Immobilization
Abstract | Full Text: HTML PDF

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Supplement 87, July 2009

Unit 17.4 Metabolic Radiolabeling of Animal Cell Glycoconjugates
Abstract | Full Text: HTML PDF

Unit 19.8 Resources for Small Regulatory RNAs
Abstract | Full Text: HTML PDF

Unit 24.6 Directed Evolution of Proteins In Vitro Using Compartmentalization in Emulsions
Abstract | Full Text: HTML PDF

Unit 25A.3 Laser Microdissection-Mediated Isolation and In Vitro Transcriptional Amplification of Plant RNA
Abstract | Full Text: HTML PDF

Appendix 6 Pathway Modulators and Inhibitors
Abstract | Full Text: HTML PDF

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Supplement 86, April 2009

Unit 3.21 DNA Cloning and Engineering by Uracil Excision
Abstract | Full Text: HTML PDF

Unit 9.5 Selection of Transfected Mammalian Cells
Abstract | Full Text: HTML PDF

Unit 19.7 Using PATIMDB to Create Bacterial Transposon Insertion Mutant Libraries
Abstract | Full Text: HTML PDF

Unit 20.7 Two-Hybrid Dual Bait System
Abstract | Full Text: HTML PDF

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Supplement 85, January 2009

Unit 10.6 Staining Proteins in Gels
Abstract | Full Text: HTML PDF

Unit 22.4 Overview of mRNA Expression Profiling Using DNA Microarrays
Abstract | Full Text: HTML PDF

Unit 22.5 Pattern Discovery in Expression Profiling Data
Abstract | Full Text: HTML PDF

Unit 23.13 Recombineering-Based Procedure for Creating Cre/loxP Conditional Knockouts in the Mouse
Abstract | Full Text: HTML PDF

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Supplement 84, October 2008

Unit 3.5 DNA-Dependent DNA Polymerases
Abstract | Full Text: HTML PDF

Unit 3.6 Template-Independent DNA Polymerases
Abstract | Full Text: HTML PDF

Unit 3.7 RNA-Dependent DNA Polymerases
Abstract | Full Text: HTML PDF

Unit 3.8 RNA Polymerases
Abstract | Full Text: HTML PDF

Unit 3.9 DNA Repair Enzymes
Abstract | Full Text: HTML PDF

Unit 3.13 Ribonucleases
Abstract | Full Text: HTML PDF

Unit 3.15 RNA Ligases
Abstract | Full Text: HTML PDF

Unit 10.7 Detection of Proteins on Blot Transfer Membranes
Abstract | Full Text: HTML PDF

Unit 14.18 Using Cell-ID 1.4 with R for Microscope-Based Cytometry
Abstract | Full Text: HTML PDF

Unit 14.19 Visualization of Microscopy-Based Spectral Imaging Data from Multi-Label Tissue Sections
Abstract | Full Text: HTML PDF

Unit 29B.5 Dietary Manipulation of Mouse Metabolism
Abstract | Full Text: HTML PDF

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Supplement 83, July 2008

Unit 10.8 Immunoblotting and Immunodetection
Abstract | Full Text: HTML PDF

Unit 11.10 Production of Monoclonal Antibody Supernatant and Ascites Fluid
Abstract | Full Text: HTML PDF

Unit 26.8 Using Morpholinos to Control Gene Expression
Abstract | Full Text: HTML PDF

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Supplement 82, April 2008

Unit 13.2 Growth and Manipulation of Yeast
Abstract | Full Text: HTML PDF

Unit 13.3B EMS and UV Mutagenesis in Yeast
Abstract | Full Text: HTML PDF

Unit 14.8 In Situ Polymerase Chain Reaction and Hybridization to Detect Low-Abundance Nucleic Acid Targets
Abstract | Full Text: HTML PDF

Unit 14.17 Using CellProfiler for Automatic Identification and Measurement of Biological Objects in Images
Abstract | Full Text: HTML PDF

Unit 20.1 Interaction Trap/Two-Hybrid System to Identify Interacting Proteins
Abstract | Full Text: HTML PDF

Unit 23.5 Production of a Heterozygous Mutant Cell Line by Homologous Recombination (Single Knockout)
Abstract | Full Text: HTML PDF

Unit 23.6 Production of a Homozygous Mutant Embryonic Stem Cell Line (Double Knockout)
Abstract | Full Text: HTML PDF

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Supplement 81, January 2008

Unit 7.1 Overview of DNA Sequencing Strategies
Abstract | Full Text: HTML PDF

Unit 9.4 Tranfection of Cultured Eukaryotic Cells Using Cationic Lipid Reagents
Abstract | Full Text: HTML PDF

Unit 11.12 Production of Polyclonal Antisera
Abstract | Full Text: HTML PDF

Unit 14.6 Immunohistochemistry
Abstract | Full Text: HTML PDF

Unit 14.11 Basic Confocal Microscopy
Abstract | Full Text: HTML PDF

Unit 26.7 Analysis of Small Endogenous RNAs
Abstract | Full Text: HTML PDF

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Supplement 80, October 2007

Unit 21.14 Chromosome Conformation Capture Carbon Copy Technology
Abstract | Full Text: HTML PDF

Unit 24.1 Overview of Receptors from Combinatorial Nucleic Acid and Protein Libraries
Abstract | Full Text: HTML PDF

Unit 25B.6 Serial Analysis of Gene Expression (SAGE): Experimental Method and Data Analysis
Abstract | Full Text: HTML PDF

Unit 29A.4 Tissue Collection for Systematic Phenotyping in the Mouse
Abstract | Full Text: HTML PDF

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Supplement 79, July 2007

Unit 1.17 E. coli Genome Manipulation by P1 Transduction
Abstract | Full Text: HTML PDF

Unit 14.7 In Situ Hybridization and Detection Using Nonisotopic Probes
Abstract | Full Text: HTML PDF

Unit 21.12 Paired-End diTagging for Transcriptome and Genome Analysis
Abstract | Full Text: HTML PDF

Unit 21.13 ChIP-chip for Genome-Wide Analysis of Protein Binding in Mammalian Cells
Abstract | Full Text: HTML PDF

Appendix 1E Radioactivity
Abstract | Full Text: HTML PDF

Appendix 1F Safe Use of Radioisotopes
Abstract | Full Text: HTML PDF

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Supplement 78, April 2007

Unit 1.16 Recombineering: Genetic Engineering in Bacteria Using Homologous Recombination
Abstract | Full Text: HTML PDF

Unit 3.17 Constructing Recombinant DNA Molecules by PCR
Abstract | Full Text: HTML PDF

Unit 4.10 Identification of Newly Transcribed RNA
Abstract | Full Text: HTML PDF

Unit 16.25 Production of Recombinant Adeno-Associated Viral Vectors for In Vitro and In Vivo Use
Abstract | Full Text: HTML PDF

Unit 29B.4 Histopathology in Mouse Metabolic Investigations
Abstract | Full Text: HTML PDF

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Supplement 77, January 2007

Unit 18.13 Isolation of Phosphopeptides by Immobilized Metal Ion Affinity Chromatography
Abstract | Full Text: HTML PDF

Unit 19.6 Analysis and Management of Microarray Gene Expression Data
Abstract | Full Text: HTML PDF

Unit 29B.3 Evaluation of Glucose Homeostasis
Abstract | Full Text: HTML PDF

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Supplement 76, October 2006

Unit 7.8 Polony DNA Sequencing
Abstract | Full Text: HTML PDF

Unit 10.1A Spectrophotometric and Colorimetric Determination of Protein Concentration
Abstract | Full Text: HTML PDF

Unit 20.5 Detection of Protein-Protein Interactions by Coprecipitation
Abstract | Full Text: HTML PDF

Unit 23.1 Overview of Gene Targeting by Homologous Recombination
Abstract | Full Text: HTML PDF

Appendix 3D Quantitation of DNA and RNA with Absorption and Fluorescence Spectroscopy
Abstract | Full Text: HTML PDF

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Supplement 75, July 2006

Unit 10.2A One-Dimensional SDS Gel Electrophoresis of Proteins
Abstract | Full Text: HTML PDF

Unit 12.1 Preparation of Nuclear and Cytoplasmic Extracts from Mammalian Cells
Abstract | Full Text: HTML PDF

Unit 21.12 Paired-End diTagging for Transcriptome and Genome Analysis
Full Text: HTML PDF

Unit 27.4 RNA Immunoprecipitation for Determining RNA-Protein Associations In Vivo
Abstract | Full Text: HTML PDF

Unit 29A.3 Collection of Blood and Plasma from the Mouse
Abstract | Full Text: HTML PDF

Unit 29B.2 Lipid and Bile Acid Analysis
Abstract | Full Text: HTML PDF

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Supplement 74, April 2006

Unit 3.20 Recombinational Cloning
Abstract | Full Text: HTML PDF

Unit 10.24 Solution Radioimmunoassay of Proteins and Peptides
Abstract | Full Text: HTML PDF

Unit 14.16 Three-Dimensional Reconstruction of Tissues
Abstract | Full Text: HTML PDF

Unit 21.11 Mapping Chromatin Interactions by Chromosome Conformation Capture
Abstract | Full Text: HTML PDF

Appendix 3F Techniques for Mammalian Cell Tissue Culture
Abstract | Full Text: HTML PDF

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Supplement 73, January 2006

Unit 15.8 High-Throughput Real-Time Quantitative Reverse Transcription PCR
Abstract | Full Text: HTML PDF

Unit 18.12 The Detection of MAPK Signaling
Abstract | Full Text: HTML PDF

Unit 29A.1 Uses of Forward and Reverse Genetics in Mice to Study Gene Function
Abstract | Full Text: HTML PDF

Unit 29A.2 Minimizing Variation Due to Genotype and Environment
Abstract | Full Text: HTML PDF

Unit 29B.1 Evaluation of Energy Homeostasis
Abstract | Full Text: HTML PDF

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Supplement 72, October 2005

Unit 10.24 Solution Radioimmunoassay of Proteins and Peptides
Full Text: HTML PDF

Unit 21.10 Identifying Chromosomal Targets of DNA-Binding Proteins by Sequence Tag Analysis of Genomic Enrichment (STAGE)
Abstract | Full Text: HTML PDF

Unit 26.4 Cloning of Small RNA Molecules
Abstract | Full Text: HTML PDF

Unit 26.6 RNAi in Transgenic Plants
Abstract | Full Text: HTML PDF

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Supplement 71, July 2005

Unit 15.8 High-Throughput Real-Time Quantitative Reverse Transcription PCR
Full Text: HTML PDF

Unit 23.10 Transgenic Mouse Colony Management
Abstract | Full Text: HTML PDF

Unit 23.11 Modification and Production of BAC Transgenes
Abstract | Full Text: HTML PDF

Unit 23.12 Regulation of Transgene Expression Using Tetracycline
Abstract | Full Text: HTML PDF

Unit 28.3 Establishment and Culture of Human Skin Fibroblasts
Abstract | Full Text: HTML PDF

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Supplement 70, April 2005

Unit 1.16 Recombineering: Genetic Engineering in Bacteria Using Homologous Recombination
Full Text: HTML PDF

Unit 19.5 Basic Protein Sequence Analysis
Abstract | Full Text: HTML PDF

Unit 21.8 Analysis of Protein Co-Occupancy by Quantitative Sequential Chromatin Immunoprecipitation
Abstract | Full Text: HTML PDF

Unit 28.1 Preparation, Culture, and Immortalization of Mouse Embryonic Fibroblasts
Abstract | Full Text: HTML PDF

Unit 28.2 Isolation and Immortalization of Lymphocytes
Abstract | Full Text: HTML PDF

Appendix 1A Common Abbreviations
Abstract |Full Text: HTML PDF

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Supplement 69, January 2005

Unit 10.23 Difference Gel Electrophoresis (DIGE) Using CyDye DIGE Fluor Minimal Dyes
Abstract | Full Text: HTML PDF

Unit 16.24 Helper-Dependent Adenoviral Vectors
Abstract | Full Text: HTML PDF

Unit 21.1 Micrococcal Nuclease Analysis of Chromatin Structure
Abstract | Full Text: HTML PDF

Unit 21.3 Chromatin Immunoprecipitation for Determining the Association of Proteins with Specific Genomic Sequences In Vivo
Abstract | Full Text: HTML PDF

Unit 22.5 Pattern Discovery in Expression Profiling Data
Full Text: HTML PDF

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Supplement 68, October 2004

Unit 2.8 Capillary Electrophoresis of DNA
Abstract | Full Text: HTML PDF

Unit 19.4 Protein Databases on the Internet
Abstract | Full Text: HTML PDF

Unit 21.8 Analysis of Protein Co-Occupancy by Quantitative Sequential Chromatin Immunoprecipitation
Full Text: HTML PDF

Unit 21.9 Defining In Vivo Targets of Nuclear Proteins by Chromatin Immunoprecipitation and Microarray Analysis
Abstract | Full Text: HTML PDF

Unit 23.9 Transgenic Mouse Production By Zygote Injection
Abstract | Full Text: HTML PDF

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Supplement 67, July 2004

Unit 4.9 Analysis of RNA by Northern and Slot Blot Hybridization
Abstract | Full Text: HTML PDF

Unit 10.4 Two-Dimensional Gel Electrophoresis
Abstract | Full Text: HTML PDF

Unit 18.12 The Detection of MAPK Signaling
Full Text: HTML PDF

Unit 22.4 Overview of mRNA Expression Profiling Using Microarrays
Full Text: HTML PDF

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Supplement 66, April 2004

Unit 10.5 Overview of Digital Electrophoresis Analysis
Abstract | Full Text: HTML PDF

Unit 10.8 Immunoblotting and Immunodetection
Full Text: HTML PDF

Unit 14.9 Whole-Mount In Situ Hybridization and Detection of RNA s in Vertebrate Embryos and Isolated Organs
Abstract | Full Text: HTML PDF

Unit 18.11 Design and Use of Analog-Sensitive Protein Kinases
Abstract | Full Text: HTML PDF

Appendix 3D Quantitation of DNA and RNA with Absorption and Fluorescence Spectroscopy
Full Text: HTML PDF

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Supplement 65, January 2004

Unit 16.9 Overview of the Baculovirus Expression System
Abstract | Full Text: HTML PDF

Unit 16.10 Maintenance of Insect Cell Cultures and Generation of Recombinant Baculoviruses
Abstract | Full Text: HTML PDF

Unit 16.11 Expression and Purification of Recombinant Proteins Using the Baculovirus System
Abstract | Full Text: HTML PDF

Unit 20.8 Interaction Trap/Two-Hybrid System to Identify Loss-of-Interaction Mutant Proteins
Abstract | Full Text: HTML PDF

Unit 21.3 Chromatin Immunoprecipitation for Determining the Association of Proteins with Specific Genomic Sequences In Vivo
Full Text: HTML PDF

Unit 26.5 RNA Interference in Cultured Drosophila Cells
Abstract | Full Text: HTML PDF

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Supplement 64, October 2003

Unit 10.7 Detection of Proteins on Blot Transfer Membranes
Full Text: HTML PDF

Unit 13.14 Overview of Schizosaccharomyces pombe
Abstract | Full Text: HTML PDF

Unit 13.15 S. pombe Strain Maintenance and Media
Abstract | Full Text: HTML PDF

Unit 13.16 Growth and Manipulation of S. pombe
Abstract | Full Text: HTML PDF

Unit 13.17 Introduction of DNA into S. pombe Cells
Abstract | Full Text: HTML PDF

Unit 26.4 Cloning of Small RNA Molecules
Full Text: HTML PDF

Appendix 5 Vectors
Abstract | Full Text: HTML PDF

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Supplement 63, July 2003

Unit 9.1 Calcium Phosphate Transfection
Abstract | Full Text: HTML PDF

Unit 10.6 Staining Proteins in Gels
Full Text: HTML PDF

Unit 14.15 Overview of Image Analysis, Image Importing, and Image Processing Using Freeware
Abstract | Full Text: HTML PDF

Unit 27.1 Agarose Gel Separation/Isolation of RNA-Protein Complexes
Abstract | Full Text: HTML PDF

Unit 27.2 Identification of RNA Binding Proteins by UV Cross-Linking
Abstract | Full Text: HTML PDF

Unit 27.3 Purification of Functional RNA-Protein Complexes Using MS 2-MBP
Abstract | Full Text: HTML PDF

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Supplement 62, April 2003

Unit 9.15 Human Somatic Cell Gene Targeting
Abstract | Full Text: HTML PDF

Unit 18.10 Use of Protein Phosphatase Inhibitors
Abstract | Full Text: HTML PDF

Unit 26.1 Overview of RNA Interference and Related Processes
Abstract | Full Text: HTML PDF

Unit 26.2 Gene Silencing by RNAi in Mammalian Cells
Abstract | Full Text: HTML PDF

Unit 26.3 RNA Interference in Caenorhabditis Elegans
Abstract | Full Text: HTML PDF

Appendix 1A Common Abbreviations
Full Text: HTML PDF

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Supplement 61, January 2003

Unit 1.16 Recombineering: Genetic Engineering in Bacteria Using Homologous Recombination
Full Text: HTML PDF

Unit 21.3 Chromatin Immunoprecipitation for Determining the Association of Proteins with Specific Genomic Sequences In Vivo

Unit 25B.6 Serial Analysis of Gene Expression (SAGE)
Full Text: HTML PDF

Unit 25B.8 Gene Expression Analysis of a Single or Few Cells
Abstract | Full Text: HTML PDF

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Supplement 60, October 2002

Unit 16.12 Transient Expression of Proteins Using COS Cells
Abstract | Full Text: HTML PDF

Unit 16.13 Expression and Purification of Epitope-Tagged Multisubunit Protein Complexes from Mammalian Cells
Abstract | Full Text: HTML PDF

Unit 16.14 Inducible Gene Expression Using an Autoregulatory, Tetracycline-Controlled System
Abstract | Full Text: HTML PDF

Unit 16.21 Overview of the HIV-1 Lentiviral Vector System
Abstract | Full Text: HTML PDF

Unit 16.22 Generation of HIV-1-Based Lentiviral Vector Particles
Abstract | Full Text: HTML PDF

Unit 16.23 Amplification Using CHO Cell Expression Vectors
Abstract | Full Text: HTML PDF

Unit 20.7 Two-Hybrid Dual Bait System
Full Text: HTML PDF

Unit 25B.7 Representational Difference Analysis
Abstract | Full Text: HTML PDF

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Supplement 59, July 2002

Unit 1.1 Media Preparation and Bacteriological Tools
Abstract | Full Text: HTML PDF

Unit 1.2 Growth in Liquid Media
Abstract | Full Text: HTML PDF

Unit 1.3 Growth on Solid Media
Abstract | Full Text: HTML PDF

Unit 1.4 Selected Topics from Classical Bacterial Genetics
Abstract | Full Text: HTML PDF

Unit 2.1A Purification and Concentration of DNA from Aqueous Solutions
Abstract | Full Text: HTML PDF

Unit 2.6 Isolation and Purification of Large DNA Restriction Fragments from Agarose Gels
Abstract | Full Text: HTML PDF

Unit 11.15 Introduction to Peptide Synthesis
Abstract | Full Text: HTML PDF

Unit 11.16 Synthetic Peptides for Production of Antibodies that Recognize Intact Proteins
Abstract | Full Text: HTML PDF

Appendix 1K Compendium of Drugs Commonly Used in Molecular Biology Research
Abstract | Full Text: HTML PDF

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Supplement 58, April 2002

Unit 4.1 Preparation of Cytoplasmic RNA from Tissue Culture Cells
Abstract | Full Text: HTML PDF

Unit 21.7 Chromatin Assembly Using Drosophila Systems
Abstract | Full Text: HTML PDF

Unit 25A.2 Preparation of Single Cells from Solid Tissues for Analysis by PCR
Abstract | Full Text: HTML PDF

Appendix 1H Safe Use of Hazardous Chemicals
Abstract | Full Text: HTML PDF

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Supplement 57, January 2002

Unit 17.6 Detection and Analysis of Proteins Modified by O-Linked N-Acetylglucosamine
Abstract | Full Text: HTML PDF

Unit 23.8 Mouse Colony Management
Abstract | Full Text: HTML PDF

Unit 25B.5 AFLP-Based Transcript Profiling
Abstract | Full Text: HTML PDF

Unit 25B.6 Serial Analysis of Gene Expression (SAGE)

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Supplement 56, October 2001

Unit 2.4 Preparation of Genomic DNA from Bacteria
Abstract | Full Text: HTML PDF

Unit 15.1 Enzymatic Amplification of DNA by PCR: Standard Procedures and Optimization
Abstract | Full Text: HTML PDF

Unit 15.2 Direct DNA Sequencing of PCR Products
Abstract | Full Text: HTML PDF

Unit 15.3 Ligation-Mediated PCR for Genomic Sequencing and Footprinting
Abstract | Full Text: HTML PDF

Unit 15.4 Molecular Cloning of PCR Products
Abstract | Full Text: HTML PDF

Unit 15.5 Enzymatic Amplification of RNA by PCR (RT-PCR)
Abstract | Full Text: HTML PDF

Unit 15.7 Quantitation of Rare DNAs by PCR
Abstract | Full Text: HTML PDF

Unit 25B.3 Differential Display of mRNA by PCR
Abstract | Full Text: HTML PDF

Unit 25B.4 Restriction-Mediated Differential Display (RMDD)
Abstract | Full Text: HTML PDF

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Supplement 55, July 2001

Unit 5.9 Construction of Bacterial Artificial Chromosome (BAC/PAC) Libraries
Abstract | Full Text: HTML PDF

Unit 12.12 Yeast One-Hybrid Screening for DNA-Protein Interactions
Abstract | Full Text: HTML PDF

Unit 14.3 In Situ Hybridization to Cellular RNA
Abstract | Full Text: HTML PDF

Unit 20.6 Identification of Protein Interactions by Far Western Analysis
Abstract | Full Text: HTML PDF

Unit 25A.1 Laser Capture Microdissection
Abstract | Full Text: HTML PDF

Unit 25B.1 Production of a Subtracted cDNA Library
Abstract | Full Text: HTML PDF

Unit 25B.2 PCR-Based Subtractive cDNA Cloning
Abstract | Full Text: HTML PDF

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Supplement 54, April 2001

Unit 10.12 HPLC of Peptides and Proteins: Preparation and System Set-Up
Abstract | Full Text: HTML PDF

Unit 10.13 HPLC of Peptides and Proteins: Standard Operating Conditions
Abstract | Full Text: HTML PDF

Unit 10.14 Reversed-Phase Isolation of Peptides
Abstract | Full Text: HTML PDF

Unit 16.22 Generation of HIV-1-Based Lentiviral Vector Particles

Unit 21.6 Assembly of Nucleosomal Templates by Salt Dialysis
Abstract | Full Text: HTML PDF

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Supplement 53, January 2001

Unit 11.18 Identification of Polyol-Responsive Monoclonal Antibodies for Use in Immunoaffinity Chromatography
Abstract | Full Text: HTML PDF

Unit 22.3 Profiling Human Gene Expression with cDNA Microarrays
Abstract | Full Text: HTML PDF

Unit 23.7 Chimeric Mouse Production by Microinjection
Abstract | Full Text: HTML PDF

Unit 24.5 Protein Selection Using mRNA Display
Abstract | Full Text: HTML PDF

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Supplement 52, October 2000

Unit 23.4 Mouse Embryonic Stem (ES) Cell Isolation
Abstract | Full Text: HTML PDF

Unit 23.5 Production of a Heterozygous Mutant Cell Line by Homologous Recombination (Single Knockout)
Full Text: HTML PDF

Unit 23.6 Production of a Homozygous Mutant Embryonic Stem Cell Line (Double Knockout)
Full Text: HTML PDF

Unit 24.2 Design, Synthesis, and Amplification of DNA Pools for Construction of Combinatorial Pools and Libraries
Full Text: HTML PDF

Unit 24.3 In Vitro Selection of RNA Aptamers to a Protein Target by Filter Immobilization
Full Text: HTML PDF

Unit 24.4 Peptide Aptamers: Dominant "Genetic" Agents for Forward and Reverse Analysis of Cellular Processes
Abstract | Full Text: HTML PDF

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Supplement 51, July 2000

Unit 2.5A Agarose Gel Electrophoresis
Abstract | Full Text: HTML PDF

Unit 2.5B Pulsed-Field Gel Electrophoresis
Abstract | Full Text: HTML PDF

Unit 8.3 Random Mutagenesis by PCR
Abstract | Full Text: HTML PDF

Unit 10.6 Staining Proteins in Gels

Unit 13.3 Genome-Wide Transposon Mutagenesis in Yeast
Abstract | Full Text: HTML PDF

Unit 19.2 Sequence Databases: Integrated Information Retrieval and Data Submission
Abstract | Full Text: HTML PDF

Unit 23.1 Overview of Gene Targeting by Homologous Recombination
Full Text: HTML PDF

Unit 23.2 Mouse Embryo Fibroblast (MEF) Feeder Cell Preparation
Abstract | Full Text: HTML PDF

Unit 23.3 Mouse Embryonic Stem (ES) Cell Culture
Abstract | Full Text: HTML PDF

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Supplement 50, April 2000

Unit 11.13 In Vitro Antibody Production
Abstract | Full Text: HTML PDF

Unit 11.14 Purification of Immunoglobulin G Fraction from Antiserum, Ascites Fluid, or Hybridoma Supernatant
Abstract | Full Text: HTML PDF

Unit 11.17 Determination of the Specific Antibody Titer
Abstract | Full Text: HTML PDF

Unit 14.14 Whole-Mount Histochemical Detection of β-Galactosidase Activity
Abstract | Full Text: HTML PDF

Unit 19.1 Internet Basics for Biologists
Abstract | Full Text: HTML PDF

Unit 19.4 Protein Databases on the Internet
Full Text: HTML PDF

Unit 21.5 Isolation of Histones and Nucleosome Cores from Mammalian Cells
Abstract | Full Text: HTML PDF

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Supplement 49, January 2000

Unit 14.12 Measurement of In Situ Hybridization
Abstract | Full Text: HTML PDF

Unit 14.13 Morphological, Biochemical, and Flow Cytometric Assays of Apoptosis
Abstract | Full Text: HTML PDF

Unit 22.1 Overview of Nucleic Acid Arrays
Abstract | Full Text: HTML PDF

Unit 22.2 Preparation of mRNA for Expression Monitoring
Abstract | Full Text: HTML PDF

Appendix 1J Common Conversion Factors
Abstract | Full Text: HTML PDF

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Supplement 48, October 1999

Unit 9.4 Transfection of Cultured Eukaryotic Cells Using Cationic Lipid Reagents
Full Text: HTML PDF

Unit 10.16 Immunoprecipitation
Abstract | Full Text: HTML PDF

Unit 17.13A Endoglycosidase and Glycoamidase Release of N-Linked Oligosaccharides
Full Text: HTML PDF

Unit 17.13B Analysis of Glycosaminoglycans with Polysaccharide Lyases
Abstract | Full Text: HTML PDF

Unit 18.9 Phosphopeptide Mapping and Identification of Phosphorylation Sites
Abstract | Full Text: HTML PDF

Unit 21.4 DN ase I and Hydroxyl Radical Characterization of Chromatin Complexes
Abstract | Full Text: HTML PDF

Appendix 3A Detection and Quantitation of Radiolabeled Proteins and DNA in Gels and Blots
Abstract | Full Text: HTML PDF

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Supplement 47, July 1999

Unit 7.0 Introduction to DNA Sequencing
Abstract | Full Text: HTML PDF

Unit 7.4A DNA Sequencing by the Dideoxy Method
Abstract | Full Text: HTML PDF

Unit 7.4B Dideoxy DNA Sequencing with Chemiluminescent Detection
Abstract | Full Text: HTML PDF

Unit 10.2A One-Dimensional SDS Gel Electrophoresis of Proteins
Full Text: HTML PDF

Unit 10.2B One-Dimensional Electrophoresis Using Nondenaturing Conditions
Abstract | Full Text: HTML PDF

Unit 21.3 Chromatin Immunoprecipitation for Determining the Association of Proteins with Specific Genomic Sequences In Vivo

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Supplement 46, April 1999

Unit 7.1 DNA Sequencing Strategies
Full Text: HTML PDF

Unit 15.1 Enzymatic Amplification of DNA by PCR: Standard Procedures and Optimization

Unit 19.3 Sequence Similarity Searching Using the BLAST Family of Programs
Abstract | Full Text: HTML PDF

Unit 20.1 Interaction Trap/Two-Hybrid System to Identify Interacting Proteins
Full Text: HTML PDF

Unit 20.5 Detection of Protein-Protein Interactions by Coprecipitation
Full Text: HTML PDF

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Supplement 45, January 1999

Unit 2.7 Separation of Small DNA Fragments by Conventional Gel Electrophoresis
Abstract | Full Text: HTML PDF

Unit 2.8 Capillary Electrophoresis of DNA
Full Text: HTML PDF

Unit 9.15 Human Somatic Cell Gene Targeting

Unit 9.16 Production of Heterozygous Mutant Cell Line

Unit 9.17 Production of Homozygous Mutant

Unit 21.1 Micrococcal Nuclease Analysis of Chromatin Structure
Full Text: HTML PDF

Unit 21.2 Separation of Histone Variants and Post-Translationally Modified Isoforms by Triton/Acetic Acid/Urea Polyacrylamide Gel Electrophoresis
Abstract | Full Text: HTML PDF

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Supplement 44, October 1998

Unit 10.9 Gel-Filtration Chromatography
Abstract | Full Text: HTML PDF

Unit 10.10 Ion-Exchange Chromatography
Abstract | Full Text: HTML PDF

Unit 10.18 Metabolic Labeling with Amino Acids
Abstract | Full Text: HTML PDF

Unit 14.10 Principles and Application of Fluorescence Microscopy
Abstract | Full Text: HTML PDF

Unit 14.11 Basic Confocal Microscopy
Full Text: HTML PDF

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Supplement 43, July 1998

Unit 10.12 HPLC of Peptides and Proteins: Preparation and System Set-Up

Unit 16.13 Expression and Purification of Epitope-Tagged Multisubunit Protein Complexes from Mammalian Cells

Unit 16.15 Overview of the Vaccinia Virus Expression System
Abstract | Full Text: HTML PDF

Unit 16.16 Preparation of Cell Cultures and Vaccinia Virus Stocks
Abstract | Full Text: HTML PDF

Unit 16.17 Generation of Recombinant Vaccinia Viruses
Abstract | Full Text: HTML PDF

Unit 16.18 Characterization of Recombinant Vaccinia Viruses and Their Products
Abstract | Full Text: HTML PDF

Unit 16.19 Gene Expression Using the Vaccinia Virus/T7 RNA Polymerase Hybrid System
Abstract | Full Text: HTML PDF

Appendix 1I Commonly Used Detergents
Abstract | Full Text: HTML PDF

Appendix 3I Statistics for the Molecular Biologist: Group Comparisons
Abstract | Full Text: HTML PDF

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Supplement 42, April 1998

Unit 2.1A Purification and Concentration of DNA from Aqueous Solutions

Unit 2.1B Purification of DNA by Anion-Exchange Chromatography
Abstract | Full Text: HTML PDF

Unit 2.2 Preparation of Genomic DNA from Mammalian Tissue
Abstract | Full Text: HTML PDF

Unit 2.11 Synthesis and Purification of Oligonucleotides
Abstract | Full Text: HTML PDF

Unit 2.12 Purification of Oligonucleotides Using Denaturing Polyacrylamide Gel Electrophoresis
Abstract | Full Text: HTML PDF

Unit 19.1 Internet Basics for Biologists

Unit 19.2 Sequence Databases: Integrated Information Retrieval and Data Submission

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Supplement 41, January 1998

Unit 1.5 Introduction to Plasmid Biology
Abstract | Full Text: HTML PDF

Unit 1.7 Large-Scale Preparation of Plasmid DNA
Abstract | Full Text: HTML PDF

Unit 10.5 Overview of Digital Electrophoresis Analysis

Unit 10.15 Purification of Recombinant Proteins and Study of Protein Interaction by Epitope Tagging
Abstract | Full Text: HTML PDF

Unit 10.22 Protein Identification and Characterization by Mass Spectrometry
Abstract | Full Text: HTML PDF

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Supplement 40, October 1997

Unit 9.2 Transfection Using DEAE-Dextran
Abstract | Full Text: HTML PDF

Unit 18.7 Assays of Protein Kinases Using Exogenous Substrates
Abstract | Full Text: HTML PDF

Unit 18.8 Permeabilization Strategies to Study Protein Phosphorylation
Abstract | Full Text: HTML PDF

Unit 20.4 Surface Plasmon Resonance for Measurements of Biological Interest
Abstract | Full Text: HTML PDF

Appendix 3G Importing Biological Materials
Abstract | Full Text: HTML PDF

Appendix 3H Kinetic Assay Methods
Abstract | Full Text: HTML PDF

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Supplement 39, July 1997

Unit 9.5 Selection of Transfected Mammalian Cells
Full Text: HTML PDF

Unit 10.8 Immunoblotting and Immunodetection

Unit 13.6 Yeast Vectors and Assays for Expression of Cloned Genes
Abstract | Full Text: HTML PDF

Unit 13.9 Manipulation of Plasmids from Yeast Cells
Abstract | Full Text: HTML PDF

Unit 13.10 Manipulation of Cloned Yeast DNA
Abstract | Full Text: HTML PDF

Unit 13.11 Preparation of Yeast DNA
Abstract | Full Text: HTML PDF

Unit 20.1 Interaction Trap/Two-Hybrid System to Identify Interacting Proteins

Unit 20.3 Phage-Based Expression Cloning to Identify Interacting Proteins
Abstract | Full Text: HTML PDF

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Supplement 38, April 1997

Unit 10.21 Overview of Peptide and Protein Analysis by Mass Spectrometry
Abstract | Full Text: HTML PDF

Unit 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

Unit 16.21 Overview of the HIV-1 Lentiviral Vector System

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Supplement 37, January 1997

Unit 1.8 Introduction of Plasmid DNA into Cells
Abstract | Full Text: HTML PDF

Unit 4.9 Analysis of RNA by Northern and Slot Blot Hybridization
Full Text: HTML PDF

Unit 8.5 Directed Mutagenesis Using the Polymerase Chain Reaction
Abstract | Full Text: HTML PDF

Unit 9.12 Large-Scale Preparation and Concentration of Retrovirus Stocks
Abstract | Full Text: HTML PDF

Unit 11.12 In Vitro Antibody Production
Full Text: HTML PDF

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Supplement 36, October 1996

Unit 4.2 Guanidine Methods for Total RNA Preparation
Abstract | Full Text: HTML PDF

Unit 9.9 Overview of the Retrovirus Transduction System
Abstract | Full Text: HTML PDF

Unit 9.10 Preparation of a Specific Retrovirus Producer Cell Line
Abstract | Full Text: HTML PDF

Unit 9.11 Transient Transfection Methods for Preparation of High-Titer Retroviral Supernatants
Abstract | Full Text: HTML PDF

Unit 9.13 Detection of Helper Virus in Retrovirus Stocks
Abstract | Full Text: HTML PDF

Unit 9.14 Retrovirus Infection of Cells In Vitro and In Vivo
Abstract | Full Text: HTML PDF

Unit 10.11A Immunoaffinity Chromatography
Abstract | Full Text: HTML PDF

Unit 10.11B Metal-Chelate Affinity Chromatography
Abstract | Full Text: HTML PDF

Unit 12.2 Mobility Shift DNA-Binding Assay Using Gel Electrophoresis
Abstract | Full Text: HTML PDF

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Supplement 35, July 1996

Unit 10.1A Spectrophotometric and Colorimetric Determination of Protein Concentration
Full Text: HTML PDF

Unit 10.1B Quantitative Amino Acid Analysis
Abstract | Full Text: HTML PDF

Unit 14.9 Whole-Mount In Situ Hybridization and Detection of RNA s in Vertebrate Embryos and Isolated Organs

Unit 18.6 Production of Antibodies That Recognize Specific Tyrosine-Phosphorylated Peptides
Abstract | Full Text: HTML PDF

Appendix 2 Commonly Used Reagents and Equipment
Abstract | Full Text: HTML PDF

Appendix 3E Introduction of Restriction Enzyme Recognition Sequences by Silent Mutation
Abstract | Full Text: HTML PDF

Appendix 3F Techniques for Mammalian Cell Tissue Culture
Full Text: HTML PDF

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Supplement 34, April 1996

Unit 5.9 Construction of Bacterial Artificial Chromosome (BAC/PAC) Libraries

Unit 5.10 Amplification of a Bacteriophage Library
Abstract | Full Text: HTML PDF

Unit 5.11 Amplification of Cosmid and Plasmid Libraries
Abstract | Full Text: HTML PDF

Unit 9.7C Use of the A. Victoria Green Fluorescent Protein to Study Protein Dynamics in Vivo
Abstract | Full Text: HTML PDF

Unit 10.20 Capillary Electrophoresis of Proteins and Peptides
Abstract | Full Text: HTML PDF

Appendix 1H Safe Use of Hazardous Chemicals

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Supplement 33, January 1996

Unit 12.11 Determination of Protein-DNA Sequence Specificity by PCR-Assisted Binding-Site Selection
Abstract | Full Text: HTML PDF

Unit 18.1 Overview of Protein Phosphorylation
Abstract | Full Text: HTML PDF

Unit 18.5 Detection of Phosphorylation by Enzymatic Techniques
Abstract | Full Text: HTML PDF

Unit 20.1 Interaction Trap/Two-Hybrid System to Identify Interacting Proteins

Unit 20.2 Affinity Purification of Proteins Binding to GST Fusion Proteins
Abstract | Full Text: HTML PDF

Appendix 1A Common Abbreviations

Appendix 1B Useful Measurements and Data
Abstract | Full Text: HTML PDF

Appendix 1C Characteristics of Amino Acids
Abstract | Full Text: HTML PDF

Appendix 1D Characteristics of Nucleic Acids
Abstract | Full Text: HTML PDF

Appendix 1E Radioactivity
Full Text: HTML PDF

Appendix 1F Safe Use of Radioisotopes
Full Text: HTML PDF

Appendix 1G Centrifuges and Rotors
Abstract | Full Text: HTML PDF

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Supplement 32, October 1995

Unit 15.7 Quantitation of Rare DNAs by PCR

Unit 17.9 Direct Chemical Analysis of Glycoconjugates for Carbohydrates
Abstract | Full Text: HTML PDF

Unit 17.11 Synthetic Glycosides as Primers of Oligosaccharide Biosynthesis and Inhibitors of Glycoprotein and Proteoglycan Assembly
Abstract | Full Text: HTML PDF

Unit 17.10B Inhibition of Glycolipid Biosynthesis
Abstract | Full Text: HTML PDF

Unit 17.14A Preparation of Glycopeptides
Abstract | Full Text: HTML PDF

Unit 17.14B Detection of Individual Glycosylation Sites on Glycoproteins
Abstract | Full Text: HTML PDF

Unit 17.16 Acid Hydrolysis for Release of Monosaccharides
Abstract | Full Text: HTML PDF

Unit 17.17A Enzymatic Release of Oligosaccharides from Glycolipids
Abstract | Full Text: HTML PDF

Unit 17.17B Endo-β-Galactosidases and Keratanase
Abstract | Full Text: HTML PDF

Unit 17.18 Analysis of Monosaccharides
Abstract | Full Text: HTML PDF

Unit 17.19A Total Compositional Analysis by High-Performance Liquid Chromatography or Gas-Liquid Chromatography
Abstract | Full Text: HTML PDF

Unit 17.19B Composition of Labeled Monosaccharides from Glycosaminoglycans
Abstract | Full Text: HTML PDF

Unit 17.20 Analysis of Oligosaccharide Negative Charge by Anion-Exchange Chromatography
Abstract | Full Text: HTML PDF

Unit 17.21A HPLC Methods for the Fractionation and Analysis of Negatively Charged Oligosaccharides and Gangliosides
Abstract | Full Text: HTML PDF

Unit 17.21B Fractionation and Analysis of Neutral Oligosaccharides by HPLC
Abstract | Full Text: HTML PDF

Unit 17.22A Nitrous Acid Degradation of Glycosaminoglycans
Abstract | Full Text: HTML PDF

Unit 17.22B Analysis of Disaccharides and Tetrasaccharides Released from Glycosaminoglycans
Abstract | Full Text: HTML PDF

Unit 17.23 Analysis of Sulfate Esters by Solvolysis or Hydrolysis
Abstract | Full Text: HTML PDF

Unit 19.3 Sequence Similarity Searching Using the BLAST Family of Programs

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Supplement 31, July 1995

Unit 3.1 Digestion of DNA with Restriction Endonucleases
Abstract | Full Text: HTML PDF

Unit 10.2 One-Dimensional SDS Gel Electrophoresis of Proteins

Unit 14.7 In situ Hybridization and Detection Using Nonisotopic Probes
Full Text: HTML PDF

Unit 14.8 In situ Polymerase Chain Reaction and Hybridization to Detect Low-Abundance Nucleic Acid Targets
Full Text: HTML PDF

Unit 17.15B β-Elimination for Release of O -GalNAc-Linked Oligosaccharides from Glycoproteins and Glycopeptides
Abstract | Full Text: HTML PDF

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Supplement 30, April 1995

Unit 7.7 Computer Manipulation of DNA and Protein Sequences
Abstract | Full Text: HTML PDF

Unit 15.1 Enzymatic Amplification of DNA by PCR: Standard Procedures and Optimization

Unit 18.1 Overview of Protein Phosphorylation

Unit 18.2 Labeling Cultured Cells with 32 P i and Preparing Cell Lysates for Immunoprecipitation
Abstract | Full Text: HTML PDF

Unit 18.3 Phosphoamino Acid Analysis
Abstract | Full Text: HTML PDF

Unit 18.4 Analysis of Phosphorylation of Unlabeled Proteins
Abstract | Full Text: HTML PDF

Unit 19.1 Internet Basics for Biologists

Unit 19.2 Sequence Databases: Information Retrieval and Data Submission

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Supplement 29, January 1995

Unit 5.5 Conversion of mRNA into Double-Stranded cDNA
Abstract | Full Text: HTML PDF

Unit 9.6 Overview of Genetic Reporter Systems
Abstract | Full Text: HTML PDF

Unit 9.7B Nonisotopic Assays for Reporter Gene Activity
Abstract | Full Text: HTML PDF

Unit 17.15A β-Elimination for Release of O -Linked Glycosaminoglycans from Proteoglycans
Abstract | Full Text: HTML PDF

Appendix 1 Safe Use of Radioisotopes

Appendix 5 Vectors

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Supplement 28, October 1994

Unit 16.1 Overview of Protein Expression in E. coli
Abstract | Full Text: HTML PDF

Unit 16.4A Introduction to Expression by Fusion Protein Vectors
Abstract | Full Text: HTML PDF

Unit 16.4B Enzymatic and Chemical Cleavage of Fusion Proteins
Abstract | Full Text: HTML PDF

Unit 16.5 Expression and Purification of lacZ and trpE Fusion Proteins
Abstract | Full Text: HTML PDF

Unit 16.6 Expression and Purification of Maltose-Binding Protein Fusions
Abstract | Full Text: HTML PDF

Unit 16.7 Expression and Purification of Glutathione- S -Transferase Fusion Proteins
Abstract | Full Text: HTML PDF

Unit 16.8 Expression and Purification of Thioredoxin Fusion Proteins
Abstract | Full Text: HTML PDF

Unit 16.10 Preparation of Insect Cell Cultures and Baculovirus Stock

Unit 16.11 Generation of Recombinant Baculoviruses and Analysis of Recombinant Protein Expression

Appendix 3A Detection and Quantitation of Radiolabeled Proteins and DNA in Gels and Blots

Appendix 3B Silanizing Glassware
Abstract | Full Text: HTML PDF

Appendix 3C Dialysis and Ultrafiltration
Abstract | Full Text: HTML PDF

Appendix 3D Quantitation of DNA and RNA with Absorption and Fluorescence Spectroscopy

Appendix 3E Introduction of Restriction Enzyme Recognition Sequences by Silent Mutation

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Supplement 27, July 1994

Unit 1.7 Large-Scale Preparation of Plasmid DNA

Unit 2.3 Preparation of Genomic DNA from Plant Tissue
Abstract | Full Text: HTML PDF

Unit 6.12 Recombination-Based Assay (RBA) for Screening Bacteriophage Lambda Libraries
Abstract | Full Text: HTML PDF

Unit 13.14 Interaction Trap/Two-Hybrid System to Identify Interacting Proteins

Unit 17.13B Analysis of Glycosaminoglycans with Polysaccharide Lyases

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Supplement 26, April 1994

Unit 2.6 Isolation and Purification of Large DNA Restriction Fragments from Agarose Gels

Unit 4.10 Identification of Newly Transcribed RNA
Full Text: HTML PDF

Unit 7.4B Dideoxy DNA Sequencing with Chemiluminescent Detection

Unit 15.8 Differential Display of mRNA by CPR

Unit 17.5 Chemical Labeling of Carbohydrates by Oxidation and Sodium Borohydride Reduction
Abstract | Full Text: HTML PDF

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Supplement 25, January 1994

Unit 2.1 Purification of DNA Using Glass Beads

Unit 2.7 Separation of Small DNA Fragments by Conventional Gel Electrophoresis

Unit 12.3 Methylation and Uracil Interference Assays for Analysis of Protein-DNA Interactions
Abstract | Full Text: HTML PDF

Unit 16.20 Expression of Proteins Using Semliki Forest Virus Vectors
Abstract | Full Text: HTML PDF

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Supplement 24, October 1993

Unit 2.14 Purification of DNA Using Anion Exchange Chromatography

Unit 3.17 Constructing Recombinant DNA Molecules by the Polymerase Chain Reaction
Full Text: HTML PDF

Unit 10.11B Metal-Chelate Affinity Chromatography

Unit 12.10 Purification of Sequence-Specific DNA-Binding Proteins by Affinity Chromatography
Abstract | Full Text: HTML PDF

Unit 13.11 Preparation of Yeast DNA

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Supplement 23, July 1993

Unit 4.9 Analysis of RNA by Northern and Slot Blot Hybridization

Unit 6.11 Use of Monoclonal Antibodies for Expression Cloning
Abstract | Full Text: HTML PDF

Unit 9.15 Overview of Gene Targeting by Homologous Recombination

Unit 9.16 Production of Heterozygous Mutant Cell Line by Homologous Recombination (Single Knockout)

Unit 9.17 Production of Homozygous Mutant Embryonic Stem Cell Line (Double Knockout)

Unit 13.12 Preparation of Yeast RNA
Abstract | Full Text: HTML PDF

Unit 17.7 Lectin Analysis of Proteins Blotted onto Filters
Abstract | Full Text: HTML PDF

Unit 17.14 Preparation of Glycopeptides

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Supplement 22, April 1993

Unit 17.1 Special Considerations for Glycoproteins and Their Purification
Abstract | Full Text: HTML PDF

Unit 17.2 Special Considerations for Proteoglycans and Glycosaminoglycans and Their Purification
Abstract | Full Text: HTML PDF

Unit 17.3 Special Considerations for Glycolipids and Their Purification
Abstract | Full Text: HTML PDF

Unit 17.4 Metabolic Radiolabeling of Animal Cell Glycoconjugates
Full Text: HTML PDF

Unit 17.6 Analysis of Saccharide Structure and Function Using Glycotransferases

Unit 17.8 Detection of Glycophospholipid Anchors on Proteins
Abstract | Full Text: HTML PDF

Unit 17.9 Phenol-Sulfuric Acid Assay for Hexoses and Pentoses

Unit 17.10 Inhibition of N-Linked Glycosylation

Unit 17.12 Sialidases
Abstract | Full Text: HTML PDF

Unit 17.13A Endoglycosidase and Glycoamidase Release of N-Linked Oligosaccharides

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Supplement 21, January 1993

Unit 2.9A Southern Blotting
Abstract | Full Text: HTML PDF

Unit 2.9B Dot and Slot Blotting of DNA
Abstract | Full Text: HTML PDF

Unit 2.10 Hybridization Analysis of DNA Blots
Abstract | Full Text: HTML PDF

Unit 7.3 Preparation of Templates for DNA Sequencing
Abstract | Full Text: HTML PDF

Unit 7.4 DNA Sequencing by the Dideoxy Method

Unit 11.11 Purification of Monoclonal Antibodies
Abstract | Full Text: HTML PDF

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Supplement 20, October 1992

Unit 3.18 Labeling and Colorimetric Detection of Nonisotopic Probes
Abstract | Full Text: HTML PDF

Unit 3.19 Chemiluminescent Detection of Nonisotopic Probes
Abstract | Full Text: HTML PDF

Unit 4.8 Primer Extension
Abstract | Full Text: HTML PDF

Unit 6.9 Overview of Strategies for Screening YAC Libraries and Analyzing YAC Clones
Abstract | Full Text: HTML PDF

Unit 6.10 Analysis of Isolated YAC Clones
Abstract | Full Text: HTML PDF

Unit 10.3 Two-Dimensional Gel Electrophoresis Using the ISO-DALT System
Abstract | Full Text: HTML PDF

Unit 15.5 Ligand-Mediated PCR for Genomic Sequencing and Footprinting

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Supplement 19, July 1992

Unit 3.1 Digestion of DNA with Restriction Endonucleases

Unit 7.7 Computer Analysis of DNA and and Protein Sequences

Unit 10.19 Isolation of Proteins for Microsequence Analysis
Abstract | Full Text: HTML PDF

Unit 13.1 Preparation of Yeast Media
Abstract | Full Text: HTML PDF

Unit 13.2 Growth and Manipulation of Yeast
Full Text: HTML PDF

Unit 15.7 Molecular Cloning of PCR Products

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Supplement 18, April 1992

Unit 5.8A Production of a Complete cDNA Library
Abstract | Full Text: HTML PDF

Unit 10.4 Two-Dimensional Gel Electrophoresis Using the O'Farrell System
Full Text: HTML PDF

Unit 10.8 Immunoblotting and Immunodetection

Unit 11.3 Isotype Determination of Antibodies
Abstract | Full Text: HTML PDF

Unit 11.4 Immunization of Mice
Abstract | Full Text: HTML PDF

Unit 11.10 Production of Monoclonal Antibody Supernatant and Ascites Fluid
Full Text: HTML PDF

Unit 13.7 Introduction of DNA into Yeast Cells
Abstract | Full Text: HTML PDF

Unit 16.14 Amplification Using CHO Cell Expression Vectors

Unit 16.19 Gene Expression Using the Vaccinia Virus/T7 RNA Polymerase Hybrid System

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Supplement 17, January 1992

Unit 7.5 DNA Sequencing by the Chemical Method
Abstract | Full Text: HTML PDF

Unit 9.10 Overview of the Retrovirus Transduction System

Unit 9.11 Preparation of a Specific Retrovirus Producer Cell Line

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

Unit 15.6 cDNA Amplification Using One-Sided (Anchored) PCR
Abstract | Full Text: HTML PDF

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Supplement 16, October 1991

Unit 7.1 DNA Sequencing Strategies

Unit 7.2 Constructing Nested Deletions for Use in DNA Sequencing
Abstract | Full Text: HTML PDF

Unit 7.3 Preparation of Templates for DNA Sequencing

Unit 7.4 DNA Sequencing by the Dideoxy Method

Unit 7.6 Denaturing Gel Electrophoresis for Sequencing
Abstract | Full Text: HTML PDF

Unit 15.2 Direct DNA Sequencing of PCR Products

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Supplement 15, July 1991

Unit 1.6 Minipreps of Plasmid DNA
Abstract | Full Text: HTML PDF

Unit 4.4 Preparation of Bacterial RNA
Abstract | Full Text: HTML PDF

Unit 8.5 Mutagenesis by the Polymerase Chain Reaction

Unit 11.2 Enzyme-Linked Immunosorbent Assays (ELISA) for Detection of Antigens
Abstract | Full Text: HTML PDF

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

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Supplement 14, April 1991

Unit 4.2 Guanidine Methods for Total RNA Preparation

Unit 9.4 Liposome-Mediated Transfection

Unit 15.5 Ligation-Mediated PCR for Genomic Sequencing and Footprinting

Unit 16.12 Overview of Protein Expression in Mammalian Cells

Unit 16.13 Transient Expression of Proteins Using COS Cells

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Supplement 13, January 1991

Unit 5.6 Ligation of Linkers or Adapters to Double-Stranded cDNA
Abstract | Full Text: HTML PDF

Unit 10.2 One-Dimensional Gel Electrophoresis of Proteins

Unit 10.6 Staining Proteins in Gels

Unit 10.18 Biosynthetic Labeling of Proteins

Unit 12.7 Detection, Purification, and Characterization of cDNA Clones Encoding DNA-Binding Proteins
Abstract | Full Text: HTML PDF

Unit 14.5 Counterstaining and Mounting of Autoradiographed In Situ Hybridization Slides
Abstract | Full Text: HTML PDF

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Supplement 12, October 1990

Unit 3.18 Nonisotopic Labeling of DNA Using Nick Translation

Unit 9.6 Reporter System Using Firefly Luciferase

Unit 13.3 Mutagenesis of Yeast Cells

Appendix 1 Characteristics of Amino Acids

Appendix 1 Characteristics of Nucleic Acids

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Supplement 11, July 1990

Unit 3.1 Digestion of DNA with Restriction Endonucleases

Unit 9.1 Calcium Phosphate Transfection

Unit 12.1 Preparation of Nuclear and Cytoplasmic Extracts from Mammalian Cells
Full Text: HTML PDF

Unit 16.1 Overview of Protein Expression in E. coli

Unit 16.2 Expression Using the T7 RNA Polymerase/Promoter System
Abstract | Full Text: HTML PDF

Unit 16.3 Expression Using Vectors with Phage Regulatory Sequences
Abstract | Full Text: HTML PDF

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Supplement 10, April 1990

Unit 1.13 Preparing Lambda DNA from Phage Lysates
Abstract | Full Text: HTML PDF

Unit 11.1 Conjugation of Enzymes to Antibodies
Abstract | Full Text: HTML PDF

Unit 11.2 Enzyme-Linked Immunosorbent Assays (ELISA) for Detection of Antigens

Unit 16.4 Introduction to Expression by Fusion Protein Vectors

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 Overview of the Baculoviral Expression System

Unit 16.9 Preparation of Insect Cell Cultures and Baculovirus Stock

Unit 16.10 Purification and Transfection of Baculoviral DNA for Generating Recombinant Viruses

Unit 16.11 Generation of Recombinant Baculoviruses and Analysis of Recombinant Protein Expression

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Supplement 9, January 1990

Unit 2.9 Electroblotting and UV Crosslinking of DNA

Unit 3.5 DNA-Dependent DNA Polymerases
Full Text: HTML PDF

Unit 4.10 Identification of Newly Transcribed RNA

Unit 9.6 Phase-Extraction Assay for CAT Activity

Unit 13.13 Preparation of Protein Extracts from Yeast
Abstract | Full Text: HTML PDF

Unit 15.1 Enzymatic Amplification of DNA by Polymerase Chain Reaction: Standard Procedures and Optimization

Unit 15.2 Direct DNA Sequencing of Polymerase Chain Reaction Products

Unit 15.3 Quantitation of Rare DNAs by the Polymerase Chain Reaction

Unit 15.4 Enzymatic Amplification of RNA by the Polymerase Chain Reaction

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Supplement 8, October 1989

Unit 1.7 Large-Scale Preparation of Plasmid DNA

Unit 1.8 Introduction of Plasmid DNA into Cells

Unit 2.11 Synthesis of Oligonucleotides

Unit 3.13 Ribonucleases
Abstract | Full Text: HTML PDF

Unit 10.17 Synthesizing Proteins In Vitro by Transcription and Translation of Cloned Genes
Abstract | Full Text: HTML PDF

Unit 12.9 Analysis of DNA-Protein Interactions Using Proteins Synthesized In Vitro from Cloned Genes
Abstract | Full Text: HTML PDF

Appendix 3 Commonly Used Biochemical Techniques

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Supplement 7, July 1989

Unit 12.4 DNase I Footprint Analysis of Protein-DNA Binding
Abstract | Full Text: HTML PDF

Unit 14.1 Fixation, Embedding, and Sectioning of Tissues, Embryos, and Single Cells
Abstract | Full Text: HTML PDF

Unit 14.2 Cryosectioning
Abstract | Full Text: HTML PDF

Unit 14.3 In Situ Hybridization to Cellular RNA

Unit 14.4 Detection of Hybridized Probe
Abstract | Full Text: HTML PDF

Unit 14.5 Counterstaining and Mounting of Autoradiographed In Situ Hybridization Slides

Unit 14.6 Immunohistochemistry
Full Text: HTML PDF

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Supplement 6, April 1989

Unit 2.9 Southern Blotting and Hybridization

Unit 2.12 Purification of Oligonucleotides Using Denaturing Polyacrylamide Gel Electrophoresis

Unit 8.2B Gene Synthesis: Assembly of Target Sequences Using Mutually Priming Long Oligonucleotides
Abstract | Full Text: HTML PDF

Unit 11.12 Production of Polyclonal Antisera in Rabbits

Unit 11.13 Purification of Immunoglobin G Fraction from Antiserum, Ascites Fluid, or Hydridoma Supernatant

Unit 11.14 Selection of an Immugenic Peptide

Unit 11.15 Production of Antipeptide Antibodies

Unit 11.16 Determination of the Specific Antibody Titer

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Supplement 5, January 1989

Unit 1.8 Introduction of Plasmid DNA into Cells

Unit 3.1 Digestion of DNA with Restriction Endonucleases

Unit 13.1 Preparation of Yeast Media

Unit 13.2 Growth and Manipulation of Yeast

Unit 13.4 Yeast Cloning Vectors and Genes
Abstract | Full Text: HTML PDF

Unit 13.6 Yeast Cloning Vectors and Genes

Unit 13.7 Introduction of DNA into Yeast Cells

Unit 13.8 Cloning Yeast Genes by Complementation
Abstract | Full Text: HTML PDF

Unit 13.9 Segregation of Plasmids from Yeast Cells

Unit 13.10 Manipulation of Cloned Yeast DNA

Unit 13.11 Preparation of Yeast DNA

Unit 13.12 Preparation of Yeast RNA

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Supplement 4, October 1988

Unit 2.5A Agarose Gel Electrophoresis

Unit 2.5B Pulsed-Field Gel Electrophoresis

Unit 2.10 Separation of Double- and Single-Stranded Nucleic Acids Using Hydroxylapatite Chromatography

Unit 3.5 DNA-Dependent DNA Polymerases

Unit 3.17 The Polymerase Chain Reaction: In Vitro Enzymatic Amplification of DNA

Unit 5.8A Production of a Completed cDNA Library

Unit 5.8B Production of a Subtracted cDNA Library

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Supplement 3, July 1988

Unit 12.2 Mobility Shift DNA-Binding Assay Using Gel Electrophoresis

Unit 12.3 Methylation Interference Assay for Analysis of DNA-Protein Interactions

Unit 12.5 UV Crosslinking of Proteins to Nucleic Acids
Abstract | Full Text: HTML PDF

Unit 12.6 Purification of DNA-Binding Proteins Using Biotin/Streptavidin Affinity Systems
Abstract | Full Text: HTML PDF

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
Abstract | Full Text: HTML PDF

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Supplement 2, April 1988

Unit 6.4 Using Synthetic Oligonucleotides as Probes
Abstract | Full Text: HTML PDF

Unit 7.3 Construction of Nested Deletions for DNA Sequencing

Unit 7.4 Sequencing by the Dideoxy Method

Unit 8.4 Linker-Scanning Mutagenesis of DNA
Abstract | Full Text: HTML PDF

Unit 10.15 High-Performance Chromatofocusing and Hydrophobic-Interaction Chromatography

Appendix 1 Standard Measurements, Data, and Abbreviations

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Supplement 1, January 1988

Unit 4.1 Preparation of Cytoplasmic RNA from Tissue Culture Cells

Unit 4.6 S1 Analysis of Messenger RNA Using Single-Stranded DNA Probes
Abstract | Full Text: HTML PDF

Unit 11.1 Conjugation of Enzymes to Antibodies

Unit 11.2 Enzyme-Linked Immunosorbent Assays (ELISA) for Detection of Antigens

Unit 11.3 Immunization of Mice

Unit 11.4 Direct Enzyme-Linked Immunosorbent Assay (ELISA) for Detection of Antibodies in Serum and Hybridoma Supernatants

Unit 11.5 Preparation of Myeloma Cells
Abstract | Full Text: HTML PDF

Unit 11.6 Preparation of Mouse Feeder Cells for Fusion and Cloning
Abstract | Full Text: HTML PDF

Unit 11.7 Fusion of Myeloma Cells with Immune Spleen Cells
Abstract | Full Text: HTML PDF

Unit 11.8 Cloning of Hybridoma Cell Lines by Limiting Dilution
Abstract | Full Text: HTML PDF

Unit 11.9 Freezing and Recovery of Hybridoma Cell Lines
Abstract | Full Text: HTML PDF

Unit 11.10 Production of Ascites Fluid

Appendix 3 Commonly Used Biochemical Techniques

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Core, 1987

Unit 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

Unit 1.5 Introduction to Plasmid Biology

Unit 1.6 Minipreps of Plasmid DNA

Unit 1.7 CsCl/Ethidium Bromide Preparations of Plasmid DNA

Unit 1.8 Introduction of Plasmid DNA into Cells

Unit 1.9 Introduction to Lambda Phages
Abstract | Full Text: HTML PDF

Unit 1.10 Lambda as a Cloning Vector
Abstract | Full Text: HTML PDF

Unit 1.11 Plating Lambda Phage to Generate Plaques
Abstract | Full Text: HTML PDF

Unit 1.12 Growing Lambda-Derived Vectors
Abstract | Full Text: HTML PDF

Unit 1.13 Making Phage DNA from Liquid Lysates

Unit 1.14 Introduction to Vectors Derived from Filamentous Phages
Abstract | Full Text: HTML PDF

Unit 1.15 Preparing and Using M13-Derived Vectors
Abstract | Full Text: HTML PDF

Unit 2.1 Purification and Concentration of DNA from Aqueous Solutions

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

Unit 2.5 Agarose Gel Electrophoresis

Unit 2.6 Isolation and Purification of Large DNA Restriction Fragments from Agarose Gels

Unit 2.7 Nondenaturing Polyacrylamide Gel Electrophoresis

Unit 2.8 Sieving Agarose Gel Electrophoresis

Unit 2.9 Southern Blotting and Hybridization

Unit 3.1 Digestion of DNA with Restriction Endonucleases

Unit 3.2 Mapping by Multiple Endonuclease Digestions
Abstract | Full Text: HTML PDF

Unit 3.3 Mapping by Partial Endonuclease Digestions
Abstract | Full Text: HTML PDF

Unit 3.4 Reagents and Radioisotopes Used to Manipulate Nucleic Acids
Abstract | Full Text: HTML PDF

Unit 3.5 DNA-Dependent DNA Polymerases

Unit 3.6 Template-Independent DNA Polymerases
Full Text: HTML PDF

Unit 3.7 RNA-Dependent DNA Polymerases
Full Text: HTML PDF

Unit 3.8 DNA-Dependent RNA Polymerases
Full Text: HTML PDF

Unit 3.9 DNA-Independent RNA Polymerases
Full Text: HTML PDF

Unit 3.10 Phosphatases and Kinases
Abstract | Full Text: HTML PDF

Unit 3.11 Exonucleases
Abstract | Full Text: HTML PDF

Unit 3.12 Endonucleases
Abstract | Full Text: HTML PDF

Unit 3.13 Nonspecific Double-Stranded Endonucleases
Full Text: HTML PDF

Unit 3.14 DNA Ligases
Abstract | Full Text: HTML PDF

Unit 3.15 RNA Ligases
Full Text: HTML PDF

Unit 3.16 Subcloning of DNA Fragments
Abstract | Full Text: HTML PDF

Unit 4.1 Preparation of Cytoplasmic RNA from Tissue Culture Cells

Unit 4.2 Guanidine Methods for Total RNA Preparation

Unit 4.3 Phenol/ SDS Method for Plant RNA Preparation
Abstract | Full Text: HTML PDF

Unit 4.4 Preparation of Bacterial RNA

Unit 4.5 Preparation of Poly(A) RNA
Abstract | Full Text: HTML PDF

Unit 4.6 S1 Analysis of Messenger RNA Using Single-Stranded DNA Probes

Unit 4.7 Ribonuclease Protection Assay
Abstract | Full Text: HTML PDF

Unit 4.8 Primer Extension

Unit 4.9 Analysis of RNA by Northern Hybridization

Unit 4.10 Identification of Newly Transcribed RNA

Unit 5.1 Genomic DNA Libraries
Abstract | Full Text: HTML PDF

Unit 5.2 cDNA Libraries
Abstract | Full Text: HTML PDF

Unit 5.3 Size Fractionation Using Sucrose Gradients
Abstract | Full Text: HTML PDF

Unit 5.4 Size Fractionation Using Agarose Gels
Abstract | Full Text: HTML PDF

Unit 5.5 Conversion of mRNA into Double-Stranded cDNA

Unit 5.6 Methylation and Addition of Linkers to Double-Stranded cDNA

Unit 5.7 Production of a Genomic Library
Abstract | Full Text: HTML PDF

Unit 5.8 Production of a cDNA Library

Unit 5.9 Amplification of a Bacteriophage Lambda Library

Unit 5.10 Amplification of Cosmid and Plasmid Libraries

Unit 6.1 Plating and Transferring Bacteriophage Libraries
Abstract | Full Text: HTML PDF

Unit 6.2 Plating and Transferring Cosmid and Plasmid Libraries
Abstract | Full Text: HTML PDF

Unit 6.3 Using DNA Fragments as Probes
Abstract | Full Text: HTML PDF

Unit 6.4 Using Synthetic Oligonucleotides as Probes

Unit 6.5 Purification of Bacteriophage Clones
Abstract | Full Text: HTML PDF

Unit 6.6 Purification of Cosmid and Plasmid Clones
Abstract | Full Text: HTML PDF

Unit 6.7 Immunoscreening of Fusion Proteins Produced in Lambda Plaques
Abstract | Full Text: HTML PDF

Unit 6.8 Immunoscreening after Hybrid Selection and Translation
Abstract | Full Text: HTML PDF

Unit 7.1 Overview of DNA Sequencing Strategies

Unit 7.2 Computer-Assisted DNA Sequence Data Entry and DNA Sequence Manipulation

Unit 7.3 Construction of Nested Deletions for DNA Sequencing Using Bal 31 Exonuclease

Unit 7.4 Sequencing by the Dideoxy Method

Unit 7.5 Sequencing by the Chemical Method

Unit 7.6 Rapid DNA Sequencing by the Chemical Degradation Method Utilizing Specialized Vectors for Cloning Nested Deletions

Unit 8.1 Oligonucleotide-Directed Mutagenesis without Phenotypic Selection
Abstract | Full Text: HTML PDF

Unit 8.2 Mutagenesis with Degenerate Oligonucleotides: Creating Numerous Mutations in a Small DNA Sequence

Unit 8.3 Region-Specific Mutagenesis

Unit 9.1 Calcium Phosphate Transfection

Unit 9.2 Transfection Using DEAE-Dextran

Unit 9.3 Transfection by Electroporation
Abstract | Full Text: HTML PDF

Unit 9.4 Optimization of Transfection

Unit 9.5 Stable Transfer of Genes into Mammalian Cells

Unit 9.6 Harvest and Assay for Chloramphenicol Acetyltransferase

Unit 9.7 Assay for Human Growth Hormone

Unit 9.8 Direct Analysis of RNA after Transfection
Abstract | Full Text: HTML PDF

Unit 9.9 Amplification Using Dihydrofolate Reductase

Unit 10.1 Colorimetric Methods

Unit 10.2 One-Dimensional Gel Electrophoresis

Unit 10.3 Two-Dimensional Gel Electrophoresis Using the ISO-DALT System

Unit 10.4 Two-Dimensional Gel Electrophoresis Using the O'Farrell System

Unit 10.5 Electroelution of Proteins from Stained Gels

Unit 10.6 Detection of Protein in Gels

Unit 10.7 Detection of Proteins on Blot Transfer Membranes

Unit 10.8 Western Blotting

Unit 10.9 Gel-Filtration Chromatography

Unit 10.10 Ion-Exchange Chromatography

Unit 10.11 Immunoaffinity Chromatography

Unit 10.12 Reversed-Phase High-Performance Liquid Chromatography

Unit 10.13 Ion-Exchange High-Performance Liquid Chromatography

Unit 10.14 Size-Exclusion High-Performance Liquid Chromatography

Unit 10.15 High-Performance Hydrophobic-Interaction Chromatography

Unit 10.16 Immunoprecipitation

Appendix 1 Standard Abbreviations and Useful Data

Appendix 2 Stock Solutions and Buffers

Appendix 3 Commonly Used Biochemical Techniques

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