Procedure for production of SV40 virus

Although this procedure is necessary for production of samples of pure virions it is usually unnecessary if a viral preparation is only required for subsequent infections. In such circumstances the cells should be harvested aseptically and processed to step e, omitting the DNase and RNase. The debris from the disrupted cells is pelleted at 15,000g for 30 min and the supernatant used as a source of virus. It should be tested for bacterial contamination with brain-heart infusion broth and Saboraud fluid medium (Appendix 4). The virus should be stored at -70°C at about 1010 p.f.u./ml. 

In 1967 Hirt developed a method of selectively extracting the small DNA of SV40 and polyoma viruses while sedimenting the larger cellular DNA. 

This may be studied in 9-cm dishes or the wells in a tissue culture 

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