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First of all, PPV can stand for Plum pox or Porcine parvovirus, I suspect you are talking about the later.
Not familiar with viral expression, but RNA of this gene should be the same as the single stranded DNA, but since you didn't do any RT, it wouldn't amplify. Only thing it can do is inhibit the reaction in the excess.
Since as I said I have no idea about this virus life cycle, it's maybe possible that the DNA undergoes some editing. Or what you see is unspecific product. If you can't get rid of it, you can try to sequence it to see what it really is.
Also Pfu is proof-reading but less eficient polymerase, you may try to amplify it with some other to see if your product is more distinct. For cloning/sequencing you can use a mix of proofredaing and normal polymerase.
And no, ssDNA can't have a product of half a length. In PCR all products become double-stranded anyway.
#151647 Problem amplifying viral gene
Posted
on 06 March 2013 - 06:54 AM
#145680 Viral genomic DNA isolation ?
Posted
on 22 November 2012 - 03:22 AM
I know Qiagen has a kit or protocol for genomic DNA extraction of this virus. Check their site.
#145684 Viral genomic DNA isolation ?
Posted
on 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.
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.
#145606 Serum inactivation:
Posted
on 20 November 2012 - 07:37 PM
