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Possible to lose a pGEM insert ? - (Apr/22/2009 )

Hi

i have a quick question, can an insert (about 500 bp) "fall out" of pGEM once ligated into it ?

am i stupid for thinking it can be lost !

cheers

V_W

-Vegemite_Warrior-

You could do an agarose gel electrophoresis, if you see a band from 3500bp you have the right one, if you see only a band from 3000, you lose it.

Or you could you cut the plasmid with EcoRI, for verify the integrity of your construction.

-Felipillo-

It can happen, depending on the type of insert. Some sequences like IRES or similar sequences can be lost.

-scolix-