A wide-ranging collection of readily reproducible methods for performing nuclear reprogramming by nuclear transfer in several different species, by fusion through both chemical treatment and electrically shocking cells, and by in vivo treatment of cells with cell extracts. Several methods of monitoring nuclear reprogramming are also presented, including the use of transgenic markers, activation of telomerase as an ES-specific marker, light and electron microscopic observation of structural changes in the nucleus, and verification of surface marker expression and the differentiation potential of stem cells. Biochemical methods are provided for the examination of chromatin protein modifications, nucleosomal footprinting, transcription factor binding, and the study of DNA methylation changes both at the specific locus level and at the level of the whole nucleus.
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Although nuclear reprogramming-the cellular process underlying cloning and the artificial generation of stem cells from somatic cells-may be considered a "black box" whose interior events are either completely unknown or at least imperfectly characterized, tantalizing clues about its operation are beginning to emerge. In Nuclear Reprogramming: Methods and Protocols, hands-on laboratory scientists describe readily reproducible methods for actually performing nuclear reprogramming by nuclear transfer in several different species, by fusion induced both chemically and electrically, and by the in vivo treatment of cells with cell extracts. Isolation of an adult stem cell type is also described. In addition, several methods of monitoring nuclear reprogramming are presented, including the use of transgenic markers, activation of telomerase as an ES-specific marker, light and electron microscopic observation of structural changes in the nucleus, and the verification of surface marker expression and differentiation potential of stem cells. Biochemical methods are provided for the examination of chromatin protein modifications, nucleosomal footprinting, transcription factor binding, and the study of DNA methylation changes both at the specific locus level and at the level of the whole nucleus. The protocols follow the successful Methods in Molecular Biology™ 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. Wide-ranging and highly practical, Nuclear Reprogramming: Methods and Protocols significantly expands the repertoire of methods available to perform, study, and understand nuclear programming, not only for cloning, but also for reprogramming somatic cells to stem cells.
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During the last half-century, nuclear transfer experiments in Xenopus have tended to fall into two categories. Read the first page Key Phrases - Statistically Improbable Phrases (SIPs): (learn more)
whole cell fusion, cloned cmhryos, enucleation pipet, hisulfite treatment, telomerasc activity, stromal stem cells, spontaneous cell fusion, injection pipet, fusion buffer, cloned bovine embryos, collecting oocytes, synthetic oviduct fluid, holding pipet, cumulus cell nuclei, somatic cell nuclei, microcell fusion, cumulus cells, somatic cell type, hepatic differentiation, brake setting, nuclear reprogramming, pluripotent embryonic stem cells, immunoprecipitated protein, unstained cells, nuclear transfer Key Phrases - Capitalized Phrases (CAPs): (learn more)
Cold Spring Harbor, Fisher Scientific, New York, Roche Molecular Biochemicals, Key Words, Molecular Probes, Alexa Fluor, Roslin Institute, Component Weight, Gene Pulser, Life Technologies, Miltenyi Biotec, Reagent Supplier Cat, Santa Cruz, Band Doctor, Becton Dickinson, Bio-Rad Laboratories, Celltracker Orange, Fuji Medical X-ray Film, Genes Der, Ivey Words, Nucleic Acids Res, Telomere Length Assay Kit New!
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Front Cover | Table of Contents | First Pages | Index | Back Cover | Surprise Me!
During the last half-century, nuclear transfer experiments in Xenopus have tended to fall into two categories. Read the first page Key Phrases - Statistically Improbable Phrases (SIPs): (learn more)
whole cell fusion, cloned cmhryos, enucleation pipet, hisulfite treatment, telomerasc activity, stromal stem cells, spontaneous cell fusion, injection pipet, fusion buffer, cloned bovine embryos, collecting oocytes, synthetic oviduct fluid, holding pipet, cumulus cell nuclei, somatic cell nuclei, microcell fusion, cumulus cells, somatic cell type, hepatic differentiation, brake setting, nuclear reprogramming, pluripotent embryonic stem cells, immunoprecipitated protein, unstained cells, nuclear transfer Key Phrases - Capitalized Phrases (CAPs): (learn more)
Cold Spring Harbor, Fisher Scientific, New York, Roche Molecular Biochemicals, Key Words, Molecular Probes, Alexa Fluor, Roslin Institute, Component Weight, Gene Pulser, Life Technologies, Miltenyi Biotec, Reagent Supplier Cat, Santa Cruz, Band Doctor, Becton Dickinson, Bio-Rad Laboratories, Celltracker Orange, Fuji Medical X-ray Film, Genes Der, Ivey Words, Nucleic Acids Res, Telomere Length Assay Kit New!
Books on Related Topics | Concordance | Text Stats Browse Sample Pages:
Front Cover | Table of Contents | First Pages | Index | Back Cover | Surprise Me!
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