(by aawara from pontiff-playground.org)
Mon Feb 25 07:17:26 EST 2008
In <1203893436.24100.0 from proxy01.news.clara.net>,
ChenHA <hzhen from freeuk.com> wrote:
> How do you know how much primers the original poster added?
The amount doesn't matter - see below.
> It is for
> the original poster to comfirm or deny, or check by runnning the gel (as
> I stated, by using the same amount), not for you to assert. Whoever
> gives you the idea the 5 or 10 picomoles is what everyone uses? I
> don't, and I have done perhaps thousands of successful PCR reactions.
> And they do fluoresces, how brightly of course depends on how much you add.
> As it happens, I see this kind of things quite often, from my own PCR.
Ethidium bromide binds single-stranded DNA extremely poorly, if at all.
It fluoresces when bound to RNA, simply because most RNA molecules do
fold-back upon themselves to form partial duplexes. Long ssDNAs also
fluoresce in the presence of ethidium for the same reason.
I find it highly improbably that your ss oligonucleotides snap back on
themselves sufficiently to fluoresce with ethidium, in the absence of
polymerization that makes them ds.
Email: echo 36434455860060025978157675027927670979097959886449930P | dc
More information about the Methods