disappearing PCR product

alvarez at mbcf.stjude.org alvarez at mbcf.stjude.org
Tue Jun 22 12:40:37 EST 1993


Hi
  I'm trying to find out a copy of the journal "PCR methods and applications"?
to identify the publisher. I heard its Cold Spring Harbor Press but I'm not
sure. If some one has access to this journal I will appreciate if you can tell
me the address to request a copy from the publisher or subscription
information. 

Thanks

A. Alvarez
alvarez at mbcf.stjude.org

  

In article <1993Jun17.102159.2504 at gnv.ifas.ufl.edu>, afc at gnv.ifas.ufl.edu writes:
> In article <1993Jun15.145544.21902 at gserv1.dl.ac.uk>, CS at abc.univie.ac.at ("Christoph Schller") writes:
>> Hi
>> We frequently (with different batches enzyme) observe the
>> disappearing of PCR products if they are EtOH prec. O/N on -20.
>> Has anybody else observed that? - And how to get rid of this
>> mysterious nuclease (phenol doesn't work).
>> Thanks
>> Christoph S.
> 
> IMHO, your problem is with precipitation, not nucleases.  No enzyme is
> going to degrade ethanol precipitated nucleic acid.
> 
> To begin with, precipitate your DNA at +4C, not -20C.  It is well 
> known (except by my graduate students) that DNA precipitates much more
> effectively at the higher temperature.
> 
> Second, centrifuge the ethanol precipitate at a higher speed or for a longer 
> time.  This is important if you have a relatively small amount of product.
> 30 minutes at 12,000 RPM is probably good.
> 
> Finally, add some carrier, either nucleic acid or protein.
> 
> Andrew Cockburn



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