{Q} Effective Inverse PCR?

Susanne Rohrer "srohrer(removethis)" at immv.unizh.ch
Wed Aug 2 02:09:42 EST 2000



Chris LaRosa wrote:

> Hyun Mi Back wrote:
> >
> > I try to clone genes using Inverse PCR.  Several times I failed. Anybody
> > could help me with successful methods or strategies?
> > Theory is simple, but it does not work yet.
> >
> > Han
> > hbkim at office.hoseo.ac.kr
>
> Query is too broad for anyone you be of use.   Try consulting a local
> expert or ask more specific questions.

Asking someone locally is probably best, but no, I don't think the question
is too broad.

I've done it before, but the thing is, it just worked the one time I knew
what I was going to get anyway. When I really went on to try and amplify
something unknown, I got a huge smear with a couple of bands but nothing
really specific. So, in theory it works, but you need really good primers
from the sequence part you know. best would be to optimize them on a plasmid
if you have one.







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