PCR ramp down rate 5°C /minute for step 3 of the protocol?

[Manickam Janakiraman]

I can't get the ramp down rate to 5°C /minute in my PCR machine. Is this step very critical or can I just leave it at the bench to cool down to 25°C  (after 95°C for 5 minutes in a heating plate).

or is there other way to get this done?

Don't want annealing step to fail.

Thanks,

Manick

[Feng Zhang]

It should be okay if you let it cool down to 25C on the bench.

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[Joyem]

    I used Applied Biosystems Veriti PCR machine, I set 3% ramp down which gave nearly 5degC/min temp down

[Manickam Janakiraman]

Thanks,

I too finally found the ramp settings in Eppendorf Mastercycler pro PCR machine and then set it at 5% ramp for 25°C step and it worked out to 5°C /minute.

Manick

[Jason Kerkvliet]

I lowered the temp in 5C steps for 1 minute each on the thermocycler and it worked fine. 

[Razieh Monjezi]

Hi Manik, 

is it like that in the second setp which is 95 for 5 min we should adjust the ramp% to 5%, or I sould add a new step with 25 and 5 min and adjust the ramp% to 5%??

Thank so much for sharing your experience.

Regards

Razieh