TA Clonning

Matthias Pfeiffer pfeiffer at thermohybaid.de
Mon Sep 10 06:38:10 EST 2001

Hi Peter,
Scott is exacly right: The insert used for cloning with the TOPO-TA should
not be 5´ phosphorylated, instead you need an OH-end. Thus, no primer
phosphorylation is required.
We used the system for routinely cloning our customised synthetic genes.
In our hands the kit worked out fine, but the amount of insert has to be
estimated with care. The ligation reaction was room temperature. Hope this
helps. Matthias


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