Posted 02 May 2010 - 02:08 AM
I'm trying to clone a PCR product into a suicide vector which would hopefully inactivate a specific gene through recombination. My vector has 2 multiple cloning sites... Does anyone know if the cloned gene orientation matters? Should it come before or after the antibiotic marker?
Posted 02 May 2010 - 05:52 AM
Posted 02 May 2010 - 10:18 PM
It shouldn't matter. How large are your recombination sites?
About 1kb. I plan to clone different restriction sites for the ends to ensure the correct orientation in the vector, but I just wasn't sure about the antibiotic marker.