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Extraction of adenovirus DNA from cell culture for PCR

Started by anyluntan, Jan 31 2005 09:42 PM

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#1 anyluntan

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Posted 31 January 2005 - 09:42 PM

Does anyone have the protocol of PCR for cell culture (adevovirus) ? Please help me. Thanks.
"the extraction of adenovirus DNA from cell culture for PCR"

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#2 labprincess

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Posted 02 February 2005 - 05:22 PM

Our lab's viral DNA (Herpesvirus) has us take several flasks of infected cells. Take S/N and spin down cell debris (1000 rpm ~5min) then take S/N (don't get any cell pellet) Spin at 12000 rpm for 2 hours. You get a little pellet (At least with herpes we do).Resuspend in a bit of buffer. Treat with SDS and Proteinase K. After that it is just a phenol chloroform extraction with ethanol precipitation. We do a lot of PCR from viral DNA and this method works for us. I don't have the protocol with me right now so this is kinda vague, but I hope that helps ya !