CLONING PROBLEMS-HELP

WS via methods%40net.bio.net (by novalidaddress from nurfuerspam.de)
Sat Oct 14 17:42:20 EST 2006


Dear Ekta,

assuming your construct is ok, then your insert might become expressed
and toxic (or is simply toxic) to your bacteria, there was some damage
to the antibiotic resistance gene or there is a problem with the
antibiotic.

You might check the antibiotic, grow at lower temperature, change the
bacterial strain and / or test other (more) clones. Did check your
cloning experiment with a restriction digest?

Maybe you want to explain us your experiment a bit more detailed?

Wolfgang


ekta jindal schrieb:

> Dear Sir/Mam
>
>   I am making a construct with pEGFP vector.I am ligating 2.7Kb isert with the vector(4.7Kb). I am getting the positive result by insert-specific primers. But i am able to isolate DNA to release the fragment to confirm the clone. The recombinant plasmid is growing very less in LB media. What could be the problem.
>   Could u please suggest me something.
>
>   Thanks
>
>
> ---------------------------------
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