Books  Animal & General Biology  Genetics 

Protocols for Gene Analysis

Handbook / Manual

Series: Methods in Molecular Biology Volume: 31

Edited By: A Harwood

416 pages, Illus

Humana Press

Spiralbound | Dec 1994 | #33065 | ISBN: 0896032582
Availability: Usually dispatched within 1 week Details
NHBS Price: £115.50 $149/€126 approx

About this book

This volume provides a set of protocols for the analysis of cloned genes. A set of techniques is described for the basic manipulation and mutagenesis of the cloned sequences. The rest of the volume is organized into a collection of techniques appropriate to the analytical route to be investigated; these include genome structure, sequence variation, gene expression, and protein function. Finally, a number of methods are described to take the step from the first gene to those with which it interacts.

...recommended...A notes section that follows each methods section may be the most useful feature. These notes are like having the author right there explaining why certain things are done or not done, what variations may be tolerated or useful, what might go wrong, and how to remedy it. - Biopharm Manufacturing


Basic Recombinant DNA Techniques:Transformation of Bacteria by Electroporation. Direct Cloning of lgt11 cDNA Inserts into a Plasmid Vector. PCR Cloning Using T- Vectors. Thermal Cycle Dideoxy DNA Sequencing. Ordered Deletions Using Exonuclease III. In Vitro Mutagenesis: 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 Link Scanning Mutations Using the PCR. Localized Random PCR Mutagenesis. Genomic Structure: The Simultaneous Isolation of RNA and DNA from Tissues and Cultured Cells. Physical Mapping of 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 2-D Electrophoresis in Agarose Gels. Inverse PCR. Sequence Variation: Use of Silver Staining to Detect Nucleic Acids. A Nonradioactive Method for the Detection of Single Strand Conformational Polymorphisms. Temperature Gradient Gel Electrophoresis for the Detection of Polymorphic DNA and RNA. TGGE in Quantitative PCR of DNA and RNA. The PGK-PCR Clonality Assay. Direct Sequencing of PCR Products. Gene Expression: The Use of Riboprobes for Analysis of Gene Expression. Quantification of Absolute Amounts of Cellular Messenger RNA by RNA-Excess Solution Hybridization. Measurements of Transcription Rates in Isolated Nuclei by Nuclear Run- Off Assay. An RNA Polymerase II In Vitro Transcription System. S1 Mapping Using Single Stranded Probes. Single Primer-Mediated PCR. 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: 6xHis-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 lgt11 Lysogen Library. Index.

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