Pharmacogenomics: Methods and Protocols

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Pharmacogenomics holds the promise of drugs tailor-made to an individual`s genetic makeup for greater efficacy and safety. In pharmacogenomics: Methods and Protocols, expert researchers and physician/clinicians describe in detail the newest and most common technologies now used in this rapidly advancing field. The authors provide readily reproducible methods for assessing the functional consequences of a certain polymorphism, evaluate the variety of genotyping platforms currently available, and discuss the management of pharmacogenomic information. Highlights include techniques for making a snapshot of the allele-specific variation in human gene expression, genome wide analysis of allele-specific expression` using oligo microarrays, in vivo assays with HaploChIP, SNP genotyping in DNA pools, and PharmGKB, the pharmacogenetics and pharmacogenomics knowledge base. The methodologies for genotyping include denaturing high-performance liquid chromatography, pyrosequencing, kinetic-fluorescence detection, mass spectrometry, and TaqMan assay for insertion/deletions. The protocols follow the successful Methods in Molecular BiologyTM series format, each offering step-by-step laboratory instructions, an introduction outlining the principles behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls. Comprehensive and highly practical, Pharmacogenomics: Methods and Protocols offers pharmacologists, geneticists, molecular biologists, and clinical chemists working in academia, biotechnology, and the pharmaceutical industry a cutting-edge collection of the key techniques used for today`s preclinical and clinical studies. Table of Contents Part I. History and Overview Pharmacogenomics:Historical Perspective and Current Status Part II. Functional Analysis of Gene Variation Transfection Assays with Allele-Specific Constructs: Functional Analysis of UDP-Glucuronosyltransferase Variants Snapshot of the Allele-Specific Variation in Human Gene Expression Genome-Wide Analysis of Allele-Specific Gene Expression Using Oligo Microarrays HaploChip: An in Vivo Assay Part III. Genotyping Techniques Aspects Influencing Genotyping Method Selection Denaturing High-Performance Liquid Chromatography for Mutation Detection and Genotyping Pyrosequencing of Clinically Relevant Polymorphisms Kinetic Fluorescence-Quenching Detection Assay for Allele Frequency Estimation Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry Fluorescence-Based Fragment Size Analysis Single-Nucleotide Polymorphism Genotyping in DNA Pools TaqMan Genotyping of Insertion/Deletion Polymorphisms Part IV. Management of Pharmacogenomic Information The Pharmacogenetics and Pharmacogenomics Knowledge Base Systems for the Management of Pharmacogenomic Information