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Genomic imprinting, the process by which the non-equivalence of the paternal and maternal genomes is established, has been fascinating us for over three decades and has provided many emerging scientists with the chance to hit their stride in a frontier posing many unexpected questions and even more surprising answers. In Genomic Imprinting: Methods and Protocols, experts in the field provide a survey of the technologies that are being applied to advance the study of imprinting. This detailed volume features new technologies that are accelerating the pace of discovery of imprinted genes and characterization of their epigenetic profile, bioinformatic procedures for prediction and comparative analyses of imprinted genes, as well as methods in embryology and basic molecular biology that have been employed for many years, some appearing in new versions for small cell numbers. Written in the highly successful Methods in Molecular Biology(TM) series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and easy to use, Genomic Imprinting: Methods and Protocols will aid scientists in unveiling both much awaited answers and all-new questions to keep this vital field busy for many exciting years to come.
List of contents
Uniparental Embryos in the Study of Genomic Imprinting.- Derivation of Induced Pluripotent Stem Cells by Retroviral Gene Transduction in Mammalian Species.- Generation of Trophoblast Stem Cells.- Immunomagnetic Purification of Murine Primordial Germ Cells.- Whole Genome Methylation Profiling by Immunoprecipitation of Methylated DNA.- Identification of Imprinted Loci by Transcriptome Sequencing.- Data Mining as a Discovery Tool for Imprinted Genes.- Engineering of Large Deletions and Duplications In Vivo.- Methylated DNA Immunoprecipitation (MeDIP) from Low Amounts of Cells.- Chromatin Immunoprecipitation to Characterize the Epigenetic Profiles of Imprinted Domains.- Quantitative Chromosome Conformation Capture.- Genome-Wide Analysis of DNA Methylation in Low Cell Numbers by Reduced Representation Bisulfite Sequencing.- Isolation of RNA and DNA from Single Preimplantation Embryos and a Small Number of Mammalian Oocytes for Imprinting Studies.- Generation of cDNA Libraries from RNP-Derived Regulatory Non-Coding RNAs.- Co-Immunoprecipitation of Long Non-Coding RNAs.- Specialized Technologies for Epigenetics in Plants.- Computational Studies of Imprinted Genes.- Insights on Imprinting from Beyond Mice and Men.- Non-Mammalian Parent-Of-Origin Effects.
Summary
Genomic imprinting, the process by which the non-equivalence of the paternal and maternal genomes is established, has been fascinating us for over three decades and has provided many emerging scientists with the chance to hit their stride in a frontier posing many unexpected questions and even more surprising answers.
