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Cell Biology Techniques

Contents

• Cell Biology Techniques
• MEDIA AND SOLUTIONS REQUIRED FOR ROUTINE ES CELL CULTURE
• Routine Culturing of ES Cells
• ISOLATION OF PRIMARY MOUSE EMBRYO FIBROBLASTS
• MITOMYCIN C TREATMENT OF PMEFs
• GAMMA IRRADIATION OF PMEF'S
• PMEF FEEDER LAYER CONCENTRATION
• ELECTROPORATION OF ES CELLS AND ISOLATION OF H/R CLONES
• KARYOTYPING ES CELLS
• VASECTOMY OF MALE MICE
• Normaski Optics
• MEDIA FOR EMBRYO CULTURE AND MANIPULATION
• Suppliers
• PREPARATION OF MICROINJECTION PIPETTES
• EMBRYO MANIPULATION PIPETTES (FLUSHERS ETC)
• COLLECTION OF MORULAE and Earlier
• Aggregation of ES cells and eight cell stage embryos
• BLASTOCYST TRANSFER
• Sending sera to testing for mouse pathogens.
• SUPEROVULATION OF MICE
• FLUSHING OUT OF BLASTOCYSTS
• Alizarin Red S (bone) and alcian blue (cartilagestaining of cleared skeletons.
• Routine Splitting and freezing of cells
• SOFT AGAR ASSAY FOR COLONY FORMATION
• VIRUS INFECTION OF FIBROBLASTS
• DNA TRANSFECTION OF EUKARYOTIC CELLS USING CALCIUM PHOSPHATE
• LIPOFECTAMINE FOR TRANSFORMATION OF EUKARYOTIC CELLS
• ELECTROPORATION OF COS-7 CELLS
• Growth and transfection of PC12 cells.
• CELL CULTURE FROM WHOLE MICE EMBRYO (DAY 10 - 11 PC)
• PCR GENOTYPING FROM the EMBRYO YOKE SACK
• b-gal staining of eukaryotic cells in vitro
• Comments on db-cAMP and forskolin
• Comments on Ro 31.8220
• IMMUNOFLUORESENCE ON CHAMBER SLIDES
• PHOTOMICROGRAPHY
• Reagents:

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