4 degree hold in PCR machine
Posted 16 April 2009 - 02:38 PM
Some people seem averse to doing overnight 4 C holds for their PCr reactions, while some don't mind. Does this really affect the PCR machine? If so, is it better to hold at a higher temperature like 10 C.
Posted 16 April 2009 - 04:16 PM
It is not necessary to hold your products at 4 deg or even 10, DNA is very stable and will last at room temperature for quite some time. The reaon people hold at a lower temperature is to limit the activity of Taq or other polymerase so as to prevent potential non-specific binding of primers and amplification from these sites. However, unless you are planning on leaving your stuff at room temp for several days, this will not be a problem, as most PCRs of 30 steps are limited by primer or dNTP availabilty.
Posted 17 April 2009 - 10:35 AM
it's a good thing that the blocks are replaceable in most cyclers.
talent does what it can
genius does what it must
i used to do what i got paid to do
Posted 27 April 2009 - 12:24 AM