I have tried to blunt a Kpn l site by Klenow fragment and ligate a
PCR fragment into the blunted site.
When I sequenced the ligation part, the site contained
GGxxxx (xxxx being PCR primer sequence) instead of Gxxxx.
Since Kpn I site is GGTAC|C,
I expected it to become single G at its 3' end after blunting with
However, that is not the case.
I have sequenced seven independent clones and got the same result.
Does this mean Kpn I site is actually GGTA|CC?
This may be a FAQ, but I could not find any bionet document from
bionet gopher search engine.
Thanks in advance for any information.
Kyoto University Faculty of Medicine