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Viral genomic DNA isolation ?

Virus DNA isolation

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3 replies to this topic

#1 @bhijit

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Posted 21 November 2012 - 10:21 PM

Hi, I am very new to virus field. I want to isolate genomic DNA of virus [parvovirus].
My friends told me there re 2 typed of protocols - 1. Involves cell lysis [freeze thaw] the use the supernatent for DNA isolation
2. Directly pellet the cultured cells and use for DNA isolation
I have genomic isolation kit for bacteria. Is it necessary for me to order new kit for Virus DNA isolation ?
Which one of the above protocol should be used ? I need the Genomic DNA for PCR ?
Thank you for your time.

#2 Curtis

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Posted 22 November 2012 - 03:22 AM

I know Qiagen has a kit or protocol for genomic DNA extraction of this virus. Check their site.

#3 leelee

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Posted 22 November 2012 - 04:59 AM

You can use either.

Are you isolating from tissue culture supernatant? Or a sample of some kind?
I probably wouldn't shell out for a kit if you are using it for PCR, unless you think you'll need really pure DNA.

#4 @bhijit

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Posted 22 November 2012 - 06:19 AM

Pardon me - but I am new to this field

Yes the virus will be from tissue culture. To recover virus [lyse the tissue culture cells and thus virus will be in supernatent] my friend suggested me to use - freeze thaw.

But he also showed me another protocol without any cell lysis. so I am confused what I should use ?

Yes as leelee suggested - PCR doesnt need pure DNA - if there are any other methods please suggest

I want to sequence this PCR before cloning so I am thinking if the DNA is pure it would be better in PCR.





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