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
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.
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