TOPO cloning

Christoph Hoefer christoph.hoefer at invitrogen.com
Fri Apr 5 06:13:00 EST 2002


Hi Daniel,
    the TOPO cloning reaction works less effeciently with fragments
over 2kb.  Prolonged incubation times more often have adverse effect
on TOPO cloning efficiency.  There is, however, a special kit for
cloning larger inserts (up to 10kb) called TOPO XL,that gives good
results.

Best regards,
Christoph
Invitrogen


danielmeb at hotmail.com ("Daniel Mebrahtu") wrote in message news:<LAW2-F162Rpy9tMTF4i0001cc14 at hotmail.com>...
> Hi all,
> 
> i am having problems with cloning a 2.5kb fragment into a c2.1 TOPO vector. 
> PCR fragments less 1Kb pop in the vector without any problem. i tried to 
> extend the incubation time to 30min. it still did not work.
> 
> do you have any idea? i called invitrogen, they couldn't give any helpful 
> suggestions.
> 
> any tip will be appreciated.
> 
> daniel
> 
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