2 replies to this topic

[Vegemite_Warrior]

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

[Felipillo]

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.

Edited by Felipillo, 26 April 2009 - 02:26 PM.

[scolix]

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