Subtractive cDNA

Sandra Pena De Ortiz spenadeo at
Thu Jul 4 10:41:12 EST 1996

On Fri, 21 Jun 1996, MARK PEGGIE wrote:

> Hi,  I wonder if someone could give me some advice.  I am about to make a 
> subtractive cDNA library but am not sure of the best approach.  I was going to 
> follow a biotin/streptavidin method but found the PCR-Select cDNA Subtraction 
> kit from Clontech.  Has anyone tried this kit - what are the advantages and 
> disadvantages.
> I'm sorry if this has been covered already.
> Cheers
> Mark Peggie


We have been constructing subtracted cDNA libraries for some time and 
have used sevearl approaches.  The best approaches have been those using 
the biotin/streptavidin steps for subtraction.  One student in our lab 
who had succesfully constructed several 
 subtracted libraries tried out the Clontech kit and it did not work.  In 
my opinion, if you carefully design a good cloning and subtraction strategy 
before starting you experiment  you have no need of using the kit and 
will get results in a reasonable amount of time.


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