Deletion after cloning - (Apr/29/2001 )

hello everybody!! I am unable to clone HIV pol gene. My insert get deleted on cloning. It becomes 300bp from 3kb, I am thankful to those who suggested me to try different host strains but it has not helped so far. Just to remind , I am using Topo Cloning kit and I have tried Topo 10 cells, Life tech max Stbl 2 cells Stratagene SURE 2 cells anf HB101 cells, but still I have the same problem. many thanks on your much anticipated suggestions.

I had a similar problem. I ended up having to use Invitrogen's pBAD topo cloning system to shut down expression. Adding glucose to the overnight culture was essential too.