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Cloning 5kb inserts

Ian A. York iayork at panix.com
Wed Jan 17 21:15:25 EST 2001

In article <3A655F8E.7598.3B007A at localhost>,
Janakiraman Vasantharajan <vasant at PLANTPATH.KSU.EDU> wrote:
>I had attempted to clone a 5kb insert (from a genomic library) into 
>the bluescript vector atleast 3 times (into Kpn I - Sma I sites) with 

In my experience both KpnI and SmaI are fussy cutters--especially KpnI.  
Are you sure it's cutting properly?  Are you sure your insert has
appropriate ends?  How have you prepared your inserts?

    Ian York   (iayork at panix.com)  <http://www.panix.com/~iayork/>
    "-but as he was a York, I am rather inclined to suppose him a
     very respectable Man." -Jane Austen, The History of England

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