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Cloning into shRNA vectors


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#1 expresson_help

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Posted 18 January 2005 - 05:54 AM

How do people get on cloning oligos for shRNA expression into shRNA expression vectors? Any tips or tricks? I've just been cloning a couple and it was very difficult!"

David
Expresson
http://www.expresson.com

#2 biotin

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Posted 18 February 2005 - 05:32 AM

Just synthesize the oligos(sense and antisense) with two overhand ends as restriction enzyme site and anneal the two oligos. Treat it as common insert to clone under polIII promoter.

Hopes this help

cheers

#3 fred_33

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Posted 01 March 2005 - 06:49 AM

hi
for improving efficency of cloning, and especially if you have compatible ends, you should order phosphorylated oligos.




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