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thermocycler - real-time versus conventional?

pcr

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#1 manta

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Posted 19 January 2013 - 04:31 AM

Hi guys,

Is there any reason why I could not use a real-time machine (standard AB7500) to run conventional PCR? We have only one regular thermocycler in the lab, and it´s currently in use. I figure the steps are the same, but maybe I´m missing something. I wouldn´t add any probes to the reaction, and of course would discard whatever wacky amplification plot I would get. It´s the first step of a nested PCR (hopefully the other group will be done before I get to the second step)...I´ll eventually run it on a gel.

Thanks!!

#2 hobglobin

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Posted 19 January 2013 - 04:53 AM

I think it should be possible, as it's more or less the same. The ramp speeds might be different depending of your conventional thermal cycler, and perhaps need adjustment.
And of course it's a matter of "wear", i.e. if your boss likes or accepts it tha the real-time machine has the usual wear when it is used, which brings it a tiny step nearer to the next routine maintenance by an expensive technician,
As we have not this machine I've no idea if you can switch of unnecessary parts such as light source and detection unit...
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#3 manta

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Posted 19 January 2013 - 05:26 AM

Thanks hobglobin...our lab space seems to have gone from 4 to 1 working thermocyclers during the past month. Hopefully this will not be routine, but IŽll give it a try today.





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