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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