TA cloning

[Wolfgang Schechinger]

Does this mean that Taq adds a dT under these conditions?
Will this work with Pwo or Pfu, too? (This would be the better 
enzymes for cloning IMHO, since they have a lower error rate)

Wolfgang

> Hi,
> You can use PCR products to TA clone them even if they were kept at
> -20. By the way no need to buy the T-clone, you can make them.
> Digest you vector (pGEM or pBSK) with EcoRV, take an aliquot and add
> to a normal PCR reaction with only dT, incubate 72 for 2 hours,
> chloroform extract, precipitate and Voila!
> 
> ..   .-. .-.   .-. .-.   .-. .-.   .-. .-.   .-. .-.   .-. .-.   .-.
> .- |\ /|||X|||\ /|||X|||\ /|||X|||\ /|||X|||\ /|||X|||\ /|||X|||\
> /|||X|| ||X|||/ \|||X|||/ \|||X|||/ \|||X|||/ \|||X|||/ \|||X|||/
> \|||X|||/ \| -' `-'   `-' `-'   `-' `-'   `-' `-'   `-' `-'   `-'
> `-'   `-' `-'   `
> 
> Mounir Izallalen			
> Recherche en Sciences de la Vie et de la Sante
> Pavillon C.E. Marchand, Laboratoire 1133
> Universite laval, Ste-Foy, PQ G1K 7P4 Canada
> Tel (418) 656-2131 ext#6785 Fax (418) 656-7176
> 
> ..   .-. .-.   .-. .-.   .-. .-.   .-. .-.   .-. .-.   .-. .-.   .-.
> .- |\ /|||X|||\ /|||X|||\ /|||X|||\ /|||X|||\ /|||X|||\ /|||X|||\
> /|||X|| ||X|||/ \|||X|||/ \|||X|||/ \|||X|||/ \|||X|||/ \|||X|||/
> \|||X|||/ \| -' `-'   `-' `-'   `-' `-'   `-' `-'   `-' `-'   `-'
> `-'   `-' `-'   `
> 
> 
> 
-----
usual disclaimers apply * This message is RNAse free - please don't touch!
-----                              
Wolfgang Schechinger         
University of Tuebingen, Germany
email: wgschech at med.uni-tuebingen.de * wwWait: http://www.medizin.uni-tuebingen.de/~wgschech/start.htm
-------