Need large amounts of pure viral dna - (Jul/16/2007 )
I am in the middle of something where I need huge copy numbers of pure viral dna (in tact viral genome), I was thinking about the different ways of getting it. Here is what I can think of so far:
1. purification from tissue culture derived plaques (not sure how to do this yet)
2. purchase premade viral stocks (actually tried it already, but the main companies out there like Boston Biomedica & Acrometrix don't carry my virus)
3. PCR? (what's the amplicon size limit for PCR? is it possible to amplify hundreds of thousands of base pairs? I have never done it for more than a few thousand bp, which is common among plasmids)
Thanks a lot for your input.
It looks like you have to make it by yourself. PCR wont work for you.
Exactly what virus are you working on? How do other lab's propagate it (if it's possible to propagate)?