Leiden Muscular Dystrophy pages - Protocols©

Adenovirus-mediated MyoD gene transfer

(by Anneke Janson en Judith van Deutekom; last modified on Feb 2004)


NOTE: initially we used a retroviral vector (see Retrovirus-mediated MyoD gene transfer)

Immunohistochemical and RNA analysis

Adenovirus-infected cell cultures are harvested for RNA-isolation and mutation detection using PTT, or fixated (in cold methanol) for immunohistochemical analysis. Using combinations of antibodies against desmin (expressed at early myogenesis), embryonic myosin and dystrophin (both expressed at later stages of myogenesis), (double-)stainings are performed to determine the transduction efficiencies, the rate of cell differentiation and the absence or presence of dystrophin (see Examples).


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