PCR purification KITS: Best one??????

Dom Spinella dspinella at chugaibio.com
Wed Aug 13 20:14:53 EST 1997


Mike Lyristis writes:
> Can anyone help me... I am at that point. I perform PCR reactions and I
> get one specific product. I have used the Qiagen clean up kit, the
> Geneclean Kit, and I have done a good ol ammonium acetate precipitation.
> What I see is conversion to what presumably is a single stranded form. I
> have repeated the purification and I get the same result consistently. I
> rang the Qiagen reps and they told me that for DNA that has some unique
> characteristic (such as high A+T content), there is some conversion to a
> single stranded form. I am having an immense amount of trouble with it.
> I am also wasting a lot of time with trouble shooting this problem which
> exists for 4 independent PCR reactions.
> 
> Has anyone any suggestions that may be of some help. I do not know what
> to do...

Well, what is it you're trying to do with your "cleaned up" PCR product?
How big is it and how are you isolating it in the fist place (prior to
clean up)?  Is it really tiny so that it is likely to melt apart under
low salt conditions?  Are you sure you have to clean it up at all?
What's your evidence that you're getting post-PCR conversion to a
single-stranded form?  A little more info would be of use.  -- D.G.
Spinella



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