Frank O. Fackelmayer
Frank.Fackelmayer at uni-konstanz.de
Tue Sep 21 09:04:14 EST 1999
Chen Linan wrote:
> What buffer I must use when annealing? And what percent of adaptor the
> method will acheive?
> Chen Linan
> >> Can two
> >> synthesized oligos can be annealed two give adaptors?
> >That´s the way, yes. Let synthesize two complementary oligos, mix equal
> >amounts, boil for 5 min, then let slowly cool to room temperature. Making
> >adaptors from custom oligos is usually cheaper, and much more flexible than
> >buying those from companies. You can, e.g. engineer an additional restriction
> >site into the adaptors that may be useful later.
You can do it in water or TE. Of course, annealing will be better in buffers
containing salt - as always in hybridization work - but annealing in TE is ok for
all routine applications. I don´t know the exact percentage of annealed oligos
you can achieve, though. It may be around 80%, perhaps higher, depending on the
sequence and length of the oligos. Anyway, it is always enough for cloning
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