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Suicide vector


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#1 fuso-girl

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Posted 02 May 2010 - 02:08 AM

Hi,

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?

Thanks !

#2 phage434

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Posted 02 May 2010 - 05:52 AM

It shouldn't matter. How large are your recombination sites?

#3 fuso-girl

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Posted 02 May 2010 - 10:18 PM

It shouldn't matter. How large are your recombination sites?


Hi,
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.




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