Protocols for Gene Analysis

Basic Recombinant Dna Techniques.- Transformation of Bacteria by Electroporation.- Direct Cloning of Agtll cDNA Inserts Into a Plasmid Vector.- PCR Cloning Using T-Vectors.- Thermal Cycle Dideoxy DNA Sequencing.- In Vitro Mutagenesis.- Ordered Deletions Using Exonuclease III.- Site-Directed Mutagenesis Using a Double-Stranded DNA Template.- Site-Directed Mutagenesis Using a Uracil-Containing Phagemid Template.- Construction of Linker-Scanning Mutations by Oligonucleotide Ligation.- Construction of Linker-Scanning Mutations Using the Polymerase Chain Reaction.- Localized Random Polymerase Chain Reaction Mutagenesis.- Genomic Structure.- The Simultaneous Isolation of RNA and DNA from Tissues and Cultured Cells.- Physical Mapping of the Human Genome by Pulsed Field Gel Electrophoresis.- Field Inversion Gel Electrophoresis.- Enhanced Chemiluminescent Detection of Horseradish Peroxidase Labeled Probes.- Nonradioactive Oligonucleotide Probe Labeling.- Analysis of DNA Restriction Enzyme Digests by Two-Dimensional Electrophoresis in Agarose Gels.- Inverse Polymerase Chain Reaction.- Sequence Variations.- Use of Silver Staining to Detect Nucleic Acids.- A Nonradioactive Method for the Detection of Single-Strand Conformational Polymorphisms (SSCP).- Temperature Gradient Gel Electrophoresis (TGGE) for the Detection of Polymorphic DNA and RNA.- TGGE in Quantitative PCR of DNA and RNA.- The PGK-PCR Clonality Assay (PPCA).- Direct Sequencing of PCR Products.- Gene Expression.- The Use of Riboprobes for the Analysis of Gene Expression.- Quantification of Absolute Amounts of Cellular Messenger RNA by RNA-Excess Solution Hybridization.- Measurements of Rate of Transcription in Isolated Nuclei by Nuclear “Run-Off” Assay.- An RNA Polymerase II In Vitro Transcription System.- SIMapping Using Single-Stranded DNA Probes.- Single Primer-Mediated Polymerase Chain Reaction.- Protein-Dna Interactions.- In Vivo DNA Footprinting by Linear Amplification.- DNA Photofootprinting with Rh(phi)2bpy3+.- The Gel Retardation Assay.- The Southwestern Assay.- Cloning DNA Binding Proteins from cDNA Expression Libraries Using Oligonucleotide Binding Site Probes.- Protein Function.- 6xffis-Ni-NTA Chromatography as a Superior Technique in Recombinant Protein Expression/Purification.- Production of 35S-Labeled Proteins in E. coli and Their Use as Molecular Probes.- Preparation and Ligand Screening of a ?gt11 Lysogen Library.