Methods Digest, Vol 49, Issue 18

Rijuta Garapaty via methods%40net.bio.net (by rijutak from googlemail.com)
Thu Jun 25 12:21:18 EST 2009


>
>
> Message: 4
> Date: Thu, 25 Jun 2009 02:42:34 GMT
> From: dk from no.email.thankstospam.net (DK)
> Subject: Re: help in PCR
> To: methods from net.bio.net
> Message-ID: <uuB0m.232$025.132 from newsfe02.iad>
>
> In article <7afsnkF1vfkkcU1 from mid.individual.net>, Peter Ellis <
> pjie2 from cam.ac.uk> wrote:
> >Rijuta Garapaty wrote:
> >>
> >> Did you vary other PCR conditions, like the primer annealing temperature
> or
> >> the number of cycles? Did you add additives like BSA to check whether
> your
> >> desired product is formed? Also, did you try varying the primer
> >> concentration in your PCR mix? While amplifying such a big fragment, the
> >> choice of polymerase is also very crucial.
> >
> >Since when was 1.4k a big fragment?
>
> It was. About 25 years ago. At least, I remember talks of  "long PCR",
> meaning 3 kbp, then. :-)
>
>
I am extremely sorry about that comment, instead of reading it as 1369 bp,
i read it as 13690 bp :(. As Peter said, 1.4 kbp fragment can very easily be
amplified using any polymerase under the standard conditions. I was
definitely wrong!


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