Hot Start PCR

Klaus.Matthaei at anu.edu.au Klaus.Matthaei at anu.edu.au
Mon Oct 5 20:32:47 EST 1992


Re Hot Start

We use an alternative.  

We use a capillary cycler where the PCR is performed in a positive
displacement pipette tip. With an automatic Pipettor the PCR mix (say 15ul)
is loaded into the capillary containing everything but the target DNA
(RNA).  The target is then loaded in a separate volume (say 5ul) so the
primers and the target don't see each other until the temperature is raised
and the volumes mix by thermal convection.  Voila, hot start without
hassle.  A second benefit is that there is no oil overlay and therefore
less mess since the tubes are sealed by the plunger.  In our hands the
method is also much more efficient than conventional cyclers.

Cheers, Klaus
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Klaus Matthaei
Gene Targeting
The John Curtin School of Medical Research
The Australian National University
E-mail: Klaus.Matthaei at anu.edu.au

"If all else fails : Read the instructions"
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