baculovirus and PCR

Allison Hanly a.hanly at cheque.uq.oz.au
Tue Mar 29 02:19:53 EST 1994


On 28 Mar 1994 04:08:31 GMT, 
Shiao Y. Wang  <sywang at whale.st.usm.edu> wrote:

>
>I have a pair of primers that I use to detect baculovirus infected shrimp.
>The problem is that the PCR reaction doesn't always work. Since
>baculovirus causes that production of occlusion bodies (protein crystals
>containing virions), one possible reason is that simply boiling the
>infected tissue in water does not release the DNA for amplification. I
>would be grateful for information on how you extract DNA from tissues for PCR.

I had the same problem when using boiled S/N to detect recombinant 
baculovirus. ( I have a hunch that boiling tended to trap the DNA in 
protein which pelleted when centrifuged after boiling) I now use the method 
detailed in Biotechniques Vol 11 pp 177-178 (1991). I have performed this 
extraction method and used it in parallel with boiled S/N. This method 
works reliably (for me) while boiling doesn't.

Admittedly I use virus in S/N but I'm sure this would work with tissue

Allison



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