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• see Cell Culture>ES cell
    

Embryo Manipulation

• Preparation of Microinjection Pipettes (PMCI)
    
• Sample preparation for microinjection (Brinkmann)
  Microinjection is the loading or transfer of a dissolved substance into a living cell. The microscopic tip of the glass microcapillary has an inner diameter between 0.2 and 1 µm [1]. This capillary is back loaded with the substance to be transferred into the cells cultured for microinjection. Typical substances include purified antibodies, DNA, RNA, peptides, or oligonucleotides.
    
• Intracytoplasmic Spermatocyte Injection (ICSI) (Brinkmann)
  Detailed technique guide for microinjection.
      
• Embryo Manipulation Pipettes (Flushers etc.) (PMCI)
    
• Micropipette Beveling (Gary A. J. Brown)
  
• ES Cell Pipette Manufacture (Gary A. J. Brown)
  Make injection needles yourself
    
• Harvesting of Cells for Blastocyst Injection (PMCI)
    
• Vasectomy of Male Mice (PMCI)
    
• Vasectomy of Mouse (Gary A. J. Brown)
  Mice are vasectomized to be mated to females used for embryo transfer. The female becomes pseudopregnant, her body geared for a pregnancy because she has been mated. Vasectomy is performed on mice at 6-8 weeks of age.
    
• Collection of Morulae and Earlier (PMCI)
    
• Aggregation of ES Cells and Eight Cell Stage Embryos (PMCI)
    
• 1-cell embryo transfer into pseudopregnant recipient female mouse (Gary A. J. Brown)
  Detailed protocol with illustration
      
• Blastocyst Transfer (PMCI)
    
• Sending Sera to Testing for Mouse Pathogens
    
• Superovulation of Mice (PMCI)
    
• Flushing Out of Blastocysts (PMCI)
    
• Alizarin Red S (Bone) and Alcian Blue (Cartilage) Staining of Cleared Skeletons  (PMCI)
    
• PCR Genotyping of Embryos and Blastocysts (PMCI)
    
• PCR Genotyping from the Embryo Yoke Sac (PMCI)
    
• ß-Gal Staining of the Murine Embryo (PMCI)
     
• b-gal Solution Assay : for quantitative analysis (Mike A. Dyer)
    
• Whole Mount X-gal Histochemistry of Transgenic Animal Tissues and Cultured Cells (Eric Mercer)
  Provides detailed procedures.
   

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