strategene's pCR-script

nishir at ohsu.edu nishir at ohsu.edu
Mon Aug 16 15:26:45 EST 1993


In article <00970F7E.B1B16D00.15079 at hal.hahnemann.edu> russos at HAL.HAHNEMANN.EDU
writes:
>Hullo,Hullo
>               Has anyone out there used the pCR-script system from Strategene
>for rapid cloning of PCR products?????? We recently tried it and got some real
>strange results.  Not to mention, we couldn't get the blue/white screening to
work. Id like to hear some opinions on the product from those who have used
it!!!
>                                                Thanx
>                                                Scott Russo
>                                                Hahnemann U., Phila
>                                                russos at hal.hahnemann.edu
> .......................................................................rave
on!!
>
We just got the pCR-script from Stratagene and also got bizarre results
(albeit, only one try so far).  What happened to you?  We tried to clone an
approximately 200bp PCR product (gel purified) and were very happy when we got
lots of white colonies.  We did about 6 minis but got only very large inserts
released.  We thought they might be concatemers, but a restriction enzyme that
should have cut the insert internally did not cut up the large insert.  So we
thought we would screen alot more colonies by PCR (using the same primers used
to generate the PCR product)... we screened 35 colonies and got only one that
produced a PCR product of the right size.  But when we grew up a mini of that
one colony, it didn't have an insert!!!  I'm still scratching my head over
this, and, frankly, I'm a little scared to go back to it again...

Any body else out there got any ideas?

Rae Nishi
OSHU
Portland OR 97201



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