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Cloning error prone PCR products


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#1 milanoj

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Posted 17 May 2002 - 05:17 AM

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.

#2 helene

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Posted 25 May 2002 - 05:50 AM

hi!

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

   good luck

   helene





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