Germ Cell Protocols [electronic resource] : Volume 2: Molecular Embryo Analysis, Live Imaging, Transgenesis, and Cloning / edited by Heide Schatten.

Totowa, NJ : Humana Press, 2004.
Methods in Molecular Biology™, 1064-3745 ; 254
Springer Protocols (Springer-12345)
Methods in Molecular Biology™, 1064-3745 ; 254
1 online resource (XIV, 418 pages)
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Germ cell research has produced enormous advances in recent years and more recently has entered into an explosive phase of new discoveries with the introduction of transgenic technologies and nuclear cloning. In Germ Cell Protocols, expert laboratory investigators describe in step-by-step detail a series of powerful techniques for the molecular and genetic analysis of germ cells in a variety of different reproductive systems. Chosen for their proven success in advancing our understanding of germ cells, these methods cover sperm and egg activation, motility, fertilization, nuclear development, nuclear cloning, the molecular characterization of specific events, the imaging of cell structures, and the cryopreservation of germ cells. Volume 2: Molecular Embryo Analysis, Live Imaging, Transgenesis, and Cloning focuses on molecular egg analysis, live egg imaging, nuclear cloning, oocyte cryopreservation, and nuclear transfer. Highlights include the identification and characterization of Oct-4, germline chromatin silencing by RNAi, cDNA subtraction and cloning in the field of trophoblast/placental development, and selective ablation. There are also methods for PCR-based cloning, differentiating embryonic stem cells into embryonic bodies, live imaging of mammalian embryos using multiphoton microscopy, and the cloning of pig, cattle, rabbit, and mouse oocytes. Each readily reproducible protocol is described in step-by-step detail and contains an introduction outlining the principle behind the technique, lists of equipment and reagents, and tips on troubleshooting and avoiding known pitfalls. An accompanying earlier volume, Sperm and Oocyte Analysis, focuses on sperm cells, oocyte analysis, oocyte maturation, fertilization, and preparation techniques. Comprehensive and cutting-edge, Germ Cell Protocols offers both novice and established researchers a gold-standard collection of hard-to-find methods for high impact research, diverse procedures that are easy-to-follow, well-illustrated, and allow a cross-species transfer of knowledge from lower vertebrates to higher mammalian systems.
Activity of the Germline-Specific Oct4-GFP Transgene in Normal and Clone Mouse Embryos
Analysis of Germline Chromatin Silencing by Double-Stranded RNA-Mediated Interference (RNAi) in Caenorhabditis elegans
cDNA Subtraction and Cloning in the Field of Trophoblast/Placental Development
PCR-Based Cloning and Differential Screening of RNAs from Xenopus Primordial Germ Cells
Differentiating Embryonic Stem Cells into Embryoid Bodies
The Derivation of Murine Embryonic Stem and Embryonic Germ Cells by Selective Ablation
Using Multiphoton Excitation to Explore the Murky Depths of Developing Embryos
Dual-Channel Confocal Ratioing of Calcium Dynamics in Living Eggs and Oocytes
A Method for Producing Cloned Pigs by Using Somatic Cells as Donors
Pronuclear Microinjection of Mouse Zygotes
Parthenogenesis and Nuclear Transfer in Rabbit Oocytes
Nuclear Transfer in Cattle
In Vitro Fertilization and Embryo Transfer in Felids
Production of Chimeric Chickens
Production of Haploid and Diploid Androgenetic Zebrafish
Construction and Detection of Fluorescent, Germline Transgenic Zebrafish
Production of Zebrafish Germline Chimeras from Cultured Cells
Studying Nuclear Reprogramming with Cell Hybrids
Cytoplasmic Transfer in Mammalian Eggs and Embryos
Conventional Technologies for Cryopreservation of Human Oocytes and Embryos
Vitrifying and Warming of Human Oocytes, Embryos, and Blastocysts
Microfluidic Removal of Cumulus Cells from Mammalian Zygotes
Mammalian Embryo Culture in a Microfluidic Device
Application of Alginate Gels to the Study of Mammalian Development
Using TUNEL in Combination with an Active Caspase-3 Immunoassay to Identify Cells Undergoing Apoptosis in Preimplantation Mammalian Embryos.
Schatten, Heide. editor.
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