Subcloning pcr products

Stefan Neuenschwander neuenschwander at inw.agrl.ethz.ch
Tue Oct 17 10:48:44 EST 1995


Having no success by sub-cloning PCR pruducts (blunt-end) I found 
this: (Lorens (1991, PCR Methods Appl 1, p140-141) a method which 
I will try as soon as i get the new primers briefly:
 1. kinasing the PCR product 
 2. filling any overhang with Klenow enzyme,
 3. form concatomer of PCR products using T4 ligase
This multimer can be efficiently cleaved by the desired
restriction endonuclease. the linear product is purified and 
ligated into the vector.
Sonds good, let me know your experience
Stefan Neuenschwander
neuenschwander at inw.agrl.ethz.ch
 




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