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