Plasmid Cloning by PCR
Type
Blog Post
...the PCR product
Isolate your PCR product from the rest of the PCR reaction using a kit, such as the QIAquick...assist with restriction enzyme digestion (usually 3-6bp)
Restriction Site: Your chosen restriction site for...plasmid of interest. You can use similar processes to add overhangs to your insert of interest for Gibson...fusing the upstream restriction site to the forward primer and the downstream restriction site to the reverse...discussed in our restriction cloning post. However, even more than with standard restriction cloning, it is...of a piece of DNA and at the same time adding restriction sites to the ends of that piece of DNA so that...itself are very similar to those outlined in our restriction cloning post with a few quirks specific to the...