Cloning 5kb inserts

[Ian A. York]

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 
-- 
    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