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Titlebook: Epigenetic Reprogramming During Mouse Embryogenesis; Methods and Protocol Katia Ancelin,Maud Borensztein Book 2021 Springer Science+Busines

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https://doi.org/10.1057/978-1-137-56652-2ming. Here we describe an optimized ChIP-seq method—STAR (Small-scale TELP-Assisted Rapid) ChIP-seq—that allows the detection of histone modifications using only a few hundred cells. This method is proven to be robust in epigenomic profiling in both embryos and cultured cells.
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Microfabricated Device for High-Resolution Imaging of Preimplantation Embryos,we describe the different steps of production and storage of the imaging device as well as its use for live imaging of mouse preimplantation embryos expressing fluorescent reporters from genetically modified alleles or after in vitro transcribed mRNA transfer by microinjection or electroporation.
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Genome-Scale CRISPR Screening for Regulators of Cell Fate Transitions,luripotency to primordial germ cell (PGC) fate, exploiting the .-GFP:.-tdTomato (SGET) mouse ESC line. The principles in this protocol can be readily adapted to characterize lineage regulators for other cell fate models and/or for other species.
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In Vitro Culture of Mouse Blastocysts to the Egg Cylinder Stage via Mural Trophectoderm Excision,hout maternal input. Here we provide a step-by-step protocol describing an experimental pipeline for isolating late blastocysts, excising (manually or via laser assistance) the mural trophectoderm, and, finally, culturing the embryo to the egg cylinder stage.
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Targeted Transgenic Mice Using CRISPR /Cas9 Technology,ation of KI animals are included. We also describe two independent CRISPR/Cas9 delivery methods to produce gene-edited animals starting from zygote-stage embryos, based either on cytoplasmic injection or electroporation.
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Studying DNA Methylation Genome-Wide by Bisulfite Sequencing from Low Amounts of DNA in Mammals,for low amounts of DNA, which include steps to estimate the minimal number of PCR cycles needed for the final library preparation to minimize PCR biases. These protocols require no more than 5 ng DNA and can easily be applied to mammalian cells available in small quantities such as early embryos or primordial germ cells.
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