Long primers for PCR

Mathias Holpert s88891 at stud-mail.uni-wuerzburg.de
Sun Nov 9 14:53:35 EST 1997


On 8 Nov 1997 05:39:56 -0800, wellcome at USERS.AFRICAONLINE.CO.KE
("Wellcome Trust, Nairobi.") wrote:

>Dear Colleagues,
>
>If there any risk of amplifying a fragment DNA with primer which are 50
>to 60 pb. Actually, I want to introduce a enzyme restriction site in
>these primers so that I can easily  detected them in agore or Acrylamide
>gel. Therefore I do need longer primers. But I wandering about doing a
>PCR such long oligonucleotides.
>My regards,

Generally, it is possible to work with primers as long as those you
intend to use. I have used primers longer than that to introduce
specific mutations into my plasmid and they worked me. But the longer
the primer, the more likely it is to form weird secondary structures
or primer-dimers. You may want to play with your PCR conditions- but
is definitely possible.

Good luck

Mathias

P.S.: Are you sure that your primers need to be this long if you just
want to introduce a new restriction site?



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