bizzare digestion problem

Elmer M. Price, Ph.D. DCRCEP at mizzou1.missouri.edu
Fri Jun 17 14:36:31 EST 1994


In article <Pine.3.89.9406161131.B14868-0100000 at unixg.ubc.ca>
brunstei at UNIXG.UBC.CA (John Brunstein) writes:
 
>
>        OK clone jockeys, here's an interesting set of observations for
>you.  Perhaps someone out there will have an idea what they mean, and
>(more importantly) a solution....
>        I have two small PCR products (184 bp and 600 bp) which are blunt
>ended. Both have restriction sites near the ends: BstEII and PvuI on the
>small one, BstEII and XbaI on the other.  Both are cloned into the SmaI
>site of pUC19, and restriction digests confirm their presence and
>orientation.
 ^^^^^^^^^^^
STUFF DELETED
>
     Subcloning problems are usually real chin scratchers.  Who knows what goes
wrong?  My only suggestion (lame as it is) is to double check the orientation
of the pUC19-600bp construct.  If in the wrong orientation, then the EcoRI and
BstE II sites will be right next to each other and a double digest may "appear"
to be linerizing the plasmid.
 
Elmer
 



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