Protocol Online logo
Top : New Forum Archives (2009-): : Molecular Cloning

High frequency of vector religation - (Feb/11/2013 )

Pages: Previous 1 2 

phage434 on Tue Feb 12 15:43:36 2013 said:

I just want to check that you are cleaning up the PCR reaction prior to cutting with the enzyme. If you don't do this, then the PCR enzyme +dNTPs will extend the DNA and trash the cut end of your insert.

Yes, I PCR amplified the 6 kb insert from another plasmid and gel purified it with a spin column prior to performing the restriction digestion.

Pages: Previous 1 2