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How to insert PreScission protease into a vector


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#1 BioExplorer

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Posted 25 February 2015 - 07:07 AM

I'm new to this, but I'm trying to insert a prescission protease into a vector via Gibson assembly. How do I design the primers when my insert is ~15bp.

Thanks

#2 labtastic

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Posted 14 March 2015 - 11:44 AM

Are you trying to fuse the entire prescion protease gene to your 15bp insert, or are you trying to fuse a precision protease cut site to your 15bp insert?

 

If the former, forget those silly expensive kits...use overlap extension pcr. 

 

If the latter, encode the cut site in a primer along with your 15bp insert. Add restriction sites to primer at both ends to ligate into vector.

 

More info would be helpful.  wacko.png






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