Cloning error prone PCR products - (May/17/2002 )
I've been experiencing some problems cloning an error-prone PCR product into a T/A vector using TOPO from Invitrogen. The products were generated using Taq so the terminal A should be present. I was thinking that the error-prone conditions may influence the addition of the terminal A residue. Does anyone have any thoughts on this? I've changed the cloning stratagy but am still very frustrated.
I've also experimented the topo TA cloning kit from invitrogen and I 've obtained poor results. Me too , I don't know the reason. But when using this kit, it's imporant using fresh PCR product because the A can be removed in a old PCR product. perhaps you should try cloning just after PCR