pcr prob

Tom Landers via methods%40net.bio.net (by galaxyglue from gmail.com)
Wed Feb 27 11:29:48 EST 2008


After reading this *very* embarrassing pissing contest, i'm amused to
point out that none of the participants examined the assumption that
EtBr was the stain used.
I always see primer bands in gels stained with "Sybr Safe".
In fact, presence or absence of a primer band in a lane with no
amplification band helps diagnose whether the failure is of the
amplicon, or of the liquid handling robot (i.e. no primers added to
that reaction).
You guys -- assuming that such pomposity and rudeness is the sole
province of male scientists -- are a real testament to the profession.

P.S. - You're still using ethidium stain? Really??



On Feb 26, 9:36 pm, "Jose de las Heras" <jose... from tiscali.co.uk> wrote:
> "ChenHA" <hz... from freeuk.com> wrote in message
>
> news:1204028525.19664.0 from proxy02.news.clara.net...
>
>
>
> > Aawara Chowdhury wrote:
> >> In <nYKwj.530$Td5.... from newsfe07.lga>,
> >>  DK <d... from no.email.thankstospam.net> wrote:
>
> >>> Interesting. I never thought about it. Will do this control next time I
> >>> PCR something. I suppose not adding template will be a good way to make
> >>> amplification inefficient :-)
>
> >> There's an old paper in Nucleic Acids Research that describes the
> >> formation of primer-dimers and a method to eliminate their formation.
>
> >> The elimination of primer-dimer accumulation in PCR, Nuc. Acids Res.
> >> 25: 3235-3241 (1995).
>
> >> AC
>
> > You think you still have any credibility left here?  Idiot.
>
> I have often noticed exactly what DK is talking about (and which sounds like
> what youa re talking about too), like him I also assumed they were unused
> primers. I also noticed that the band is not always present, and the amount
> of DNA synthesised is not large enough to account for the removal of all the
> primers... But I never thought too much about it, and just carried on with
> the job at hand.
> What Aawara says makes sense to me, and because I'm curious, I'll run the
> reaction without Taq just to see.
> There's no need to get personal about this sort of thing.
>
> About credibility...
> I think the person who resorts to personal insults stands a higher chance to
> lose it, rather than the person who merely disagrees about an explanation
> and provides arguments to support his view. But then, maybe I'm an idiot
> too.
>
> Jose




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