Thermocycler for RAPDS

Prasit Palitpolkanpim - GRSO grppl at MUCC.MAHIDOL.AC.TH
Thu Feb 24 18:21:43 EST 1994


Dear Dr. Samac,
	I once used Techne PHC-2 thermocycler for RAPD work.  It worked 
fine. I encountered two problems though.  First, the temperature may not 
be homogeneous across the heat block (my time setting is about 30 sec at 
each stage).  The second is the PHC-2 start timing when the temperature 
is 2oC from the set temp.  If the tap water used to cool the heat block 
is not cold enough, this may mean that the set annealing temp may never 
be reached.
	I am not sure these are relevant.
	Regards.
	Prasit P.

On 24 Feb 1994, Deborah A. Samac wrote:

> Dear Netters,
> 
> We have been attempting to do RAPD PCR using a Techne PHC-3 thermocycler and had
> very poor results; we get only a few faint bands or no bands at all.  However, 
> duplicate reactions run in a Coy Tempcycler II down the hall give reproducible, 
> bright bands.  Both machines were set to run the same profiles of 1 min 94 C, 1 
> minute at 36 C, 2 minutes 72 C for 40 cycles.  We've tried adjusting the ramping
> of the Techne to match the slower temperature changes of the Coy but this did 
> not improve amplification.  The temperture settings on the Techne also had to be
> adjusted to match the temperatures attained by the Coy.  For example, although 
> the Techne was set for 94 C, it only reached 94 degrees for about 5 seconds 
> before starting the next step.  We're about ready to pitch the Techne (from a 
> very high window) and purchase a new machine.  Any advice on getting the Techne 
> to work will be appreciated. (I have not been able to get any technical support 
> on this problem from Techne.) 
> 
> Thanks,
> Debby
> 
> Deborah A. Samac
> Department of Plant Pathology, 495 Borlaug Hall
> University of Minnesota
> St. Paul, MN   55108
> (612) 625-1243 (office) (612) 625-9728 (fax)
> 
> 
> 
> 



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