Capillary Air Thermal Cyclers

Peter Kuhlman pkuhlman at bio.indiana.edu
Thu Jul 22 11:41:29 EST 1993


In article <1993Jul20.074548.26744 at gserv1.dl.ac.uk> jclewley at crc.ac.uk (Dr. J.P. Clewley) writes:
 >Does anyone have any experience of using the Corbett Research FTS-IS
 >capillary thermal cycler, or the Idaho technology ATC? Are there
 >particular contamination problems with the pipette tip sealing setup?
 >Problems with leakage from the  capillaries (evaporation) during
 >cycling? Apart from speed would you buy one in preference to a
 >conevntional (0.5 ml tube) machine? Any other comments about these
 >types of machine. They're superficially appealling because of their
 >speed - particularly in a diagnostic virology lab.
 >I don't have the address or fax or email # of Idaho Technology,
 >or their UK distributer. Can anyone supply it?

People in our lab use both conventional and capillary PCR (Idaho Tech),
and it seems that with a few exceptions, the capillary machine is at least
as effective in terms of yield and purity of product as the Cetus or MJ
machines.  And of course, it is MUCH faster -- one can routinely perform
and analyze three sequential PCR experiments in one day.  That, combined
with the small reaction volume, means that one can do a reaction to optimize
conditions and follow it with a scaled up version for cloning or sequencing
and still have time to gel-purify the PCR product in _one_ day's work.

That said, I'd like to stress two things:
1)  Members of the lab spent a fair amount of time optimizing reaction
conditions and reagents for the capillary machine before most of us
came to use and love it.

2)  In spite of that optimization, some of us have been unable to get
certain template/primer combinations to amplify adequately (or at all) 
on the capillary machine but have used a conventional PCR machine with 
excellent results.  So it's not black and white.  (is anything?... =)  )

As regards tube sealing and contamination concerns, we haven't had any
problems with the former -- three seconds in a bunsen burner flame works
every time -- or the latter -- we work with fairly highly conserved primers
paired with template DNAs from eubacteria, plants, animals, and fungi,
and have not had much cross-contamination trouble at all -- but I have often
wondered that contamination isn't more of a problem...

Finally, if it helps, when we recently made the decision to purchase a
second machine, we chose to supplement the Idaho Tech with a conventional
design, for flexibility's sake.


-Peter Kuhlman
pkuhlman at bio.indiana.edu
Palmer Laboratory
Indiana University



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