Phosphorylation of Pfu PCR-products

Zhiyogn Zhao zxz4 at PSU.EDU
Fri May 9 11:40:33 EST 1997

> Someone stated recently that it is necessary to phosphorylate Pfu PCR
> products prior to e.g. EcoRV subcloning. Is this really necessary?

No, it is not. Although the PCR products have no 5'phosphate groups, the 
vector has (if you do not dephosphorylated it). To avoid vector 
self-ligation, just simply add some EcoRV in the ligation reaction.

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